Automated system and method for processing multiple liquid-based specimens

ABSTRACT

An automated system and method for individually processing multiple specimens of particulate matter-containing liquid in respective containers. The containers are transported seriatim along a processing path to present them to, at least, a preprocessing apparatus (e.g., a mixing head), and then to a specimen acquisition apparatus, which removes preprocessed specimen fluid from the container for subsequent analytical testing or evaluation. Each apparatus is actuated in response to presentation of a container thereto so as to carry out its respective operation independently. An exemplary system can include, in sequential order, stations for container loading and unloading, container uncapping and cap disposal, specimen mixing, filter loading, specimen acquisition (e.g., by aspiration, and then slide printing), filter disposal, and container resealing. The system is capable of unattended operation for many hours at a time, and can interface with an integrated data management system to provide fully integrated specimen and information management in a complete diagnostic laboratory system.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims the benefit of commonly owned U.S. provisionalapplication Nos. 60/330,092, filed Oct. 19, 2001, 60/372,080, filed Apr.15, 2002, and 60/373,658, filed Apr. 19, 2002, all of which areincorporated herein by reference. This application also is related tocommonly owned U.S. non-provisional application Ser. No. 10/122,151,filed Apr. 15, 2002, which is also incorporated herein by reference.

TECHNICAL FIELD

The present disclosure is directed to apparatus and methods forcollecting and processing specimens of particulate matter-containingliquid, e.g., biological fluid, including collecting and depositing ontoa microscope slide or other surface a uniform layer of particulatestherefrom (e.g., cells) suitable for examination (e.g., use in cytologyprotocols).

BACKGROUND ART

Diagnostic cytology, particularly in the area of clinical pathology,bases cytological interpretations and diagnoses on examination of cellsand other microscopic objects. The accuracy of the screening process anddiagnosis, and the preparation of optimally interpretable samples fromspecimens typically depends upon adequate specimen and samplepreparation. In this regard the ideal sample would consist of amonolayer of substantially evenly spaced cells, which enablescytotechnologists, cytopathologists, other medical professionals, andautomated screening and diagnostic equipment to view or image the cellsmore clearly so that abnormalities can be identified more readily, moreaccurately and more reproducibly. Newer methodologies such asimmunocytochemistry and cytometric image analysis require preparationapparatus and methods that are safe, effective, accurate, precise,reproducible, inexpensive, efficient, fast and convenient.

Cytological examination of a sample begins with obtaining specimensincluding a sample of cells from the patient, which can typically bedone by scraping, swabbing or brushing an area, as in the case ofcervical specimens, or by collecting body fluids, such as those obtainedfrom the chest cavity, bladder, or spinal column, or by fine needleaspiration or fine needle biopsy. In a conventional manual cytologicalpreparation, the cells in the fluid are then transferred directly or bycentrifugation-based processing steps onto a glass microscope slide forviewing. In a typical automated cytological preparation, a filterassembly is placed in the liquid suspension and the filter assembly bothdisperses the cells and captures the cells on the filter. The filter isthen removed and placed in contact with a microscope slide. In all ofthese endeavors, a limiting factor in the sample preparation protocol isadequately separating solid matter from its fluid carrier, and in easilyand efficiently collecting and concentrating the solid matter in a formreadily accessible to examination under a microscope.

Currently, biological specimens are collected for cytologicalexaminations using special containers. These containers usually containa preservative and transport solution for preserving the cytologyspecimen during shipment from the collection site to the diagnosticcytology laboratory. Further, cytology specimens collected from the bodycavities using a swab, spatula or brush are also preserved in specialcontainers with fixatives (e.g., alcohol or acetone fixatives) prior totransferring cells onto the slide or membrane for staining orexamination. Specimen containers are known that allow a liquid-basedbiological specimen to be processed directly in the container so as toobtain a substantially uniform layer of cells on a collection site (in afilter housing defining a particulate matter separation chamber) that isassociated with the container itself. See, for example, U.S. Pat. Nos.5,301,685; 5,471,994; 6,296,764; and 6,309,362, of Raouf A. Guirguis,all of which are incorporated herein by reference.

The filtration techniques taught in these patents in practice haveyielded fairly good results in terms of obtaining close to a monolayerof cells on slides, but there is room for improvement. Further, thetypes of specimen containers disclosed in these patents requirespecially configured apertured covers and adapters therefor that aredesigned to mate with the filter housing, and with suction equipment(e.g., a syringe or a mechanized vacuum source) used to aspirate liquidfrom the container and draw it through the filter. In addition,extraction of the filter so that it can be pressed against a microscopeslide to transfer collected cells to the slide requires disassembly ofthe cooperating parts of the cover and/or adapters associated therewith.If the processing is done by automated equipment, special handlingdevices are required to carry out such disassembly. All of thiscomplexity adds time, and material and labor cost to the processingrequired prior to the actual cytology examination.

In general, automated equipment thus far developed for processingliquid-based specimens have not performed with sufficient consistency,reliability, speed and automation to satisfy current and projected needsin cancer screening and other cytology-based medical, analytical,screening and diagnostic procedures. The vial-based automated processingsystem disclosed herein provides a safe, elegant and effective solutionto these problems.

SUMMARY DISCLOSURE OF THE INVENTION

The specimen vial disclosed herein houses a complete processingassembly, typically one for mixing the liquid-based specimen therein andfor holding a filter on which a uniform layer of cells can be collectedfrom the specimen. It is expected that the specimen vial would beprepackaged with a liquid preservative solution, as is commonplace, andsent to the point-of-care site for specimen collection.

The processing assembly is coupled to a simple cover for the vial bymeans of a simple and inexpensive releasable coupling. When the cover isremoved at the point-of-care site (physician's office, clinic, hospital,etc.), the processing assembly remains with the cover to allow medicalpersonnel easy access to the container interior for insertion of abiological specimen into the vial. The cover, along with the attachedprocessing assembly, is then replaced to seal the vial. The vial maythen be sent to a laboratory for processing.

When the vial is manipulated in a simple way while still closed, theprocessing assembly detaches from the cover and remains in the vial foraccess by automated or manual laboratory equipment when the cover issubsequently removed. In a preferred embodiment, a downward force on thecenter of the cover is all that is required to detach the processingassembly from the cover. In contrast with the prior art specimen vialsdiscussed above, the vial of the present invention requires no furtherinteraction with the cover, which can be removed by a simple uncappingdevice and is discarded to avoid contamination. Ribs inside the vialsupport the processing assembly in the proper position for access duringprocessing. This self-contained vial and processing assembly arrangementminimizes human operator exposure to biohazards, such as tuberculosis orother pathogens in sputum or in other specimens types, such as urine,spinal tap fluid, gastric washings, fine-needle aspirates, andgynecological samples.

The automated specimen processing apparatus disclosed herein is referredto as the “LBP” device (for liquid-based preparation), and is designedto produce slides of high quality and consistency. The LBP device alsocan be interfaced with a device for detecting and/or quantifyingmultiple morphologic, cytochemical, and/or molecular changes at thecellular level.

During the past two years or so, a review of the literature andreanalysis of existing data have led to the identification of a panel ofmolecular diagnostic reagents that are capable of detecting andcharacterizing lung cancer, which is the most common cancer, with highsensitivity and specificity. See, for instance, commonly owned U.S.patent application Ser. Nos. 10/095,297 and 10/095,298, both filed Mar.12, 2002, and Ser. No. 10/241,753, filed Sep. 12, 2002. Here, the cellscan be reacted with antibodies and or nucleic-acid “probes” thatidentify a pattern of changes that is consistent with a diagnosis ofcancer. The molecular system can utilize algorithms fine tuned for thattumor heterogeneity.

Identifying molecular changes at the cellular level is one of the wayscancer can be detected early and at a more curable stage. Such moleculardiagnostic devices can be used for early detection and diagnosis withthe necessary sensitivity and specificity to justify their use aspopulation-based screens for individuals who are at-risk for developingcancer. Such a molecular diagnostic device also can be used tocharacterize the tumor, thereby permitting the oncologist to stratifyhis/her patients, to customize therapy, and to monitor patients in orderto assess therapeutic efficacy and disease regression, progression orrecurrence. The availability of such tests will also foster thedevelopment of new and more effective therapeutic approaches for thetreatment of early stage disease.

Such molecular diagnostics are designed to balance cost and testperformance. While screening tests must exhibit high sensitivity andspecificity, cost is always a critical factor, as the tests aretypically directed to performing on a large number of individuals who,while at-risk, do not typically have symptomatic evidence of thedisease. In this respect, the present LBP device can be interfaced witha molecular diagnostic device to develop a system for automaticallydiagnosing cancer, with a minimum or no human intervention.Alternatively, the present LBP device can be interfaced with a pathologywork station, where medical professionals can observe individual slidesprepared by the LBP device. The resulting diagnosing system, regardlesswhether an automated device or a manual observation device isinterfaced, can be interfaced with an integrated data management systembased on specialized software and a computer operating system to managedata entry and exchange of information, and network with the laboratoryand hospital information systems.

The present LBP device transports multiple specimen vials of the noveltype mentioned above sequentially through various processing stationsand produces fixed specimens on slides, each slide being bar-coded andlinked through a data management system to the vial and the patient fromwhich it came. Fresh slides are automatically removed one at a time froma cassette, and each is returned to the same cassette after a specimenis fixed thereon. Multiple slide cassettes can be loaded into the LBPdevice, and the device will automatically draw fresh slides from thenext cassette after all of the slides of the preceding one have beenused. The slide cassettes preferably are configured for liquid immersionand interfacing with automated staining equipment that will stain thespecimens without having to remove the slides from the cassette. In thisregard the cassettes preferably have slots that allow for liquiddrainage, and slots or other means that cooperate with the hooksnormally used in the staining equipment to suspend other types of slideholders. The same slide cassettes are also configured to interface withautomated diagnostic equipment and other devices that are part of anintegrated system.

While specimen vials can be loaded into the transport manually, the fullbenefits of automation can be realized by using an optional vialhandling system that automatically loads specimen vials for processing,and removes each one after its processing is complete. In one example ofsuch a handling system the vials initially are loaded manually intospecial space-saving trays that hold up to forty-one vials each. Up toeight trays can be loaded into the LBP device, and the device willprocess all of them sequentially, removing one at a time from a tray andreturning processed (and resealed) vials to a tray. The trays also canbe used for storing and retrieving processed vials.

Each vial is transported through the LBP device on a computer-controlledconveyor, in its own receptacle. (In the example disclosed the conveyorhas thirty receptacles.) The vials and the receptacles are keyed so thatthe vials proceed along the processing path in the proper orientation,and cannot rotate independently of its respective receptacle. They firstpass a bar code reader (at a data acquisition station), where the vialbar code is read, and then proceed stepwise through the followingprocessing stations of the LBP device: an uncapping station including acap disposal operation; a primary mixing or dispersal station; a filterloading station; a specimen acquisition and filter disposal station; acell deposition station; and a re-capping station. There is also a slidepresentation station, at which a fresh microscope slide is presented tothe specimen acquisition station for transfer of the specimen from thefilter to the slide. Each of the stations operates independently on thevial presented to it by the conveyor. The conveyor will not advanceuntil all of the operating stations have completed their respectivetasks, except that the conveyor may be allowed to advance after thefilter, with the adhered specimen, is moved to begin specimen transferto the slide.

The vial uncapping station has a rotary gripper that unscrews the coverfrom the vial, and discards it. Before doing so, however, the uncappinghead presses on the center of the cover to detach the internalprocessing assembly from the cover. The primary mixing station has anexpanding collet that grips the processing assembly, lifts it slightlyand moves (e.g., spins) it in accordance with a specimen-specificstirring protocol (speed and duration). The filter loading stationdispenses a specimen-specific filter type into a particulate matterseparation chamber (manifold) at the top of the processing assembly. Thespecimen acquisition station has a suction head that seals to the filterat the top of the processing assembly and first moves the processingassembly slowly to re-suspend particulate matter in the liquid-basedspecimen. Then the suction head draws a vacuum on the filter to aspiratethe liquid-based specimen from the vial and past the filter, leaving amonolayer of cells on the bottom surface of the filter. Thereafter themonolayer specimen is transferred to a fresh slide, and the vial movesto the re-capping station, where a foil seal is applied to the vial.

An improved filter system ensures that the highest quality monolayerspecimens are produced. Specimen liquid flows through the filter as wellas substantially across the front surface of the filter. Specifically,the specimen liquid is made to have a secondary flow component acrossthe filter surface. The secondary flow is designed to flow radiallyoutwardly or have a substantial radial component, which creates ashearing action that flushes or washes clusters of relatively weaklyadhering particulates so that a more uniformly distributed and thinnerlayer can be formed on the front surface of the filter. In this respect,the present system includes a peripheral outlet through which specimenliquid can flow from the area adjacent the front surface of the filter.

The filter assembly preferably has a holder, a frit seated in theholder, and a membrane filter positioned over and in contact with theouter surface of the frit. The frit can extend beyond the end of theholder. The membrane filter can be attached to the holder. The sidewallportion extending beyond the holder forms an area through which thespecimen liquid can flow, creating a secondary flow. The holder can beconfigured so that the frit is slightly bowed outwardly at the center sothat when pressure is applied to a slide during the specimentransferring step, the central portion of the frit flattens to moreevenly contact the membrane filter to the slide for more effectivetransfer.

The manifold at the upper end of the processing assembly seats thefilter assembly with the membrane filter side facing down. The manifoldpreferably has a substantially conically configured bottom wall thatrises from the central inlet (which communicates with the dependingsuction tube portion of the processing assembly). The filter assemblyand the conically configured bottom wall form a manifold chamber thathas a slight gap at its periphery, forming a peripheral outlet, byvirtue of raised members or standoffs that act as spacers. The standoffscan have channels between them through which the specimen liquid canflow out of the manifold chamber.

Various preferred materials and possible alternatives are specifiedherein for several components of the system. It is to be understood thatmaterial choices are not limited to the specific materials mentioned,and that the choice of an alternate material is governed by manyfactors, among them functionality, molding accuracy, durability,chemical resistance, shelf life, cost, availability, and/or opticalclarity (e.g., to address user requirements or marketing issues).

In its most basic aspect the invention claimed herein is directed to anautomated method and automated apparatus for individually processingmultiple specimens of particulate matter-containing liquid in respectivecontainers. The containers are transported seriatim along a processingpath to present them to, at least, a preprocessing apparatus adapted topreprocess the specimen fluid in any container presented to it, and thento a specimen acquisition apparatus adapted to remove preprocessedspecimen fluid from any container presented to it for subsequentanalytical testing or evaluation. Each apparatus is actuated in responseto presentation of a container thereto so as to carry out its respectiveoperation independently. The specimens may contain biological material.

The preprocessing apparatus may act on the specimen to disperseparticulate components of the specimen fluid, e.g., by mixing, while thespecimen acquisition apparatus may collect a sample of the particulates,e.g., on a filter. In the case of filtration, a filter loading headupstream of the specimen acquisition apparatus dispenses a filter into acontainer-borne particulate matter separation chamber, the filterloading head operating independently of the other apparatus in responseto any container presented thereto. The filter-borne sample may betransferred to a slide.

According to other aspects of the invention, the method may involveadditional operations, including container uncapping prior to specimenpreprocessing, and recapping of containers after processing is complete.The apparatus similarly may include additional heads or other devicesfor performing these operations. Each operation is carried outindependently in response to presentation of container to the respectiveoperating station.

BRIEF DESCRIPTION OF THE DRAWING FIGURES

Preferred embodiments of the disclosed system and the invention,including the best mode for carrying out the invention, are described indetail below, purely by way of example, with reference to theaccompanying drawing, in which:

FIG. 1 is a vertical sectional view through a specimen vial for use withthe LBP device, showing the processing assembly (stirrer) in the vialcoupled to the cover;

FIG. 2 a is a front elevational view of the container portion of thevial;

FIG. 2 b is a top plan view of the container, shown with the stirrerremoved;

FIG. 3 is a top plan view of the stirrer;

FIG. 4 is a bottom plan view of the liner that fits within the cover;

FIG. 5 is an exploded vertical sectional view of the stirrer and afilter assembly adapted for use in the stirrer;

FIG. 6 is a vertical sectional view of the upper portion of the stirrer,showing the filter assembly in place in the particulate matterseparation chamber;

FIG. 7 a is a partial schematic view of the arrangement depicted in FIG.6, showing the flow of liquid and particulate matter separatedtherefrom;

FIG. 7 b is a view similar to FIG. 7 a, showing liquid flow in a priorart filter system;

FIG. 8 is an exploded, cross-sectional view of the filter assembly;

FIG. 9 is a schematic illustration of the dimensional configuration ofthe flow manifold;

FIG. 10 is a vertical sectional view of the specimen vial similar toFIG. 1, but showing the stirrer detached from the cover;

FIG. 10 a is a partial vertical sectional view similar to FIG. 10,showing a modification of the stirrer;

FIG. 11 is a top plan view of the LBP device;

FIG. 11 a is a schematic diagram of the operating sequence of the LBPdevice;

FIG. 12 is a front perspective view of the LBP device, with certainparts removed for clarity;

FIG. 13 is a rear perspective view of a portion of the LBP device,showing the auto loader/unloader mechanism;

FIG. 14 is a top plan view of the auto loader/unloader mechanism;

FIG. 15 is a front elevational view of the auto loader/unloadermechanism;

FIG. 15 a is a detail sectional view taken along line 15 a-15 a in FIG.14;

FIG. 16 is an elevational view of an alternative embodiment of a gripperfor the auto loader/unloader mechanism;

FIG. 17 is a perspective view of a specimen vial tray used in the autoloader/unloader mechanism;

FIG. 18 is an enlarged detail view taken at encircling line 18 in FIG.17;

FIG. 19 is a bottom perspective view of the specimen vial tray of FIG.17;

FIG. 20 is a perspective view of three stacked specimen vial trays;

FIG. 21 is a block diagram showing specimen vial handling and data flow;

FIG. 21 a is a pictorial diagram showing an overall laboratory systemincorporating the LBP device;

FIG. 21 b is a relational database table;

FIG. 22 is a block diagram showing a computer or work station;

FIG. 23 is a facsimile of a computer screen;

FIG. 24 is a facsimile of another computer screen;

FIG. 25 is a facsimile of two computer screens;

FIG. 26 is a vertical sectional view of a specimen vial being uncapped;

FIG. 27 is a front elevational view, partly in section, of a specimenvial engaged by the uncapping head of the LBP device;

FIG. 28 is a top plan view of the uncapping head, taken along line 28-28in FIG. 27;

FIG. 29 is a side elevational view of the uncapping station of the LBPdevice;

FIG. 30 is a sectional view taken along line 30-30 in FIG. 29;

FIG. 31 is a top plan view of the uncapping station of FIG. 29;

FIG. 32 is a vertical sectional view of a specimen container showingengagement by the primary stirring head;

FIG. 33 is a side elevational view of the primary stirring station ofthe LBP device;

FIG. 34 is a front elevational view of the primary stirring station;

FIG. 35 is a top plan view of the primary stirring station;

FIG. 36 is a vertical sectional view of a specimen container duringfilter loading;

FIG. 37 is a side elevational view of the magazine portion of the filterloading station of the LBP device;

FIG. 38 is a front elevational view of the pusher portion of the filterloading station;

FIG. 39 is a top plan view of the pusher portion of the filter loadingstation;

FIG. 40 is a top plan view of the magazine portion of the filter loadingstation;

FIG. 41 is a vertical sectional view of a specimen container duringspecimen acquisition;

FIG. 42 is a vertical sectional view of a specimen container duringspecimen transfer to a slide;

FIG. 43 is a side elevational view of the specimen acquisition stationof the LBP device;

FIG. 44 is a front elevational view of the lower portion of the specimenacquisition station;

FIG. 45 is a top plan view of the specimen acquisition station, partlyin section, taken along line 45-45 in FIG. 43;

FIG. 46 is a top plan view of the specimen acquisition station;

FIG. 47 is a schematic of a bubble flow meter used in the specimenacquisition station;

FIG. 47 a is a schematic of a modification of the flow meter of FIG. 47;

FIG. 48 is a schematic of a vacuum system used in the specimenacquisition station;

FIG. 49 is an operation chart for the vacuum system of FIG. 48;

FIG. 50 is a front perspective view of the re-capping station of the LBPdevice;

FIG. 51 is a side elevational view of the re-capping station;

FIG. 52 is a front perspective view of a slide cassette used in the LBPdevice;

FIG. 53 is a detail perspective view of the slide cassette taken fromFIG. 52;

FIG. 54 is a rear perspective view of the slide cassette;

FIG. 55 is a side elevational view of the slide cassette;

FIG. 56 is a top plan view of the slide presentation system of the LBPdevice; and

FIG. 57 is a side elevational view of the slide presentation system.

DETAILED DESCRIPTION OF BEST MODE

A full description of this vial-based specimen handling and processingsystem must begin with the vial itself, which consists of a container, acover and a processing assembly (stirrer) in the vial.

Specimen Vial

Referring to FIGS. 1, 2 a and 2 b, the vial 10 comprises a container 20,a cover 30 and a processing assembly 40. Processing assembly 40 isdesigned to carry out several functions, among them mixing, and for thispreferred rotary embodiment will be referred to as a stirrer for thesake of convenience. Container 20 preferably is molded of a translucentplastic, preferably polypropylene, and has a substantially cylindricalwall 21, surrounding its longitudinal axis, joined to a conical bottomwall 22. Possible alternative plastics include ABS andpolycyclohexylenedimethylene terephthalate, glycol (commerciallyavailable from Eastman Kodak Co. under the name EASTAR® DN004). A smallportion 24 of wall 21 preferably is flat, the outer surface of the flatportion adapted to receive indicia, e.g., a bar code label, containinginformation concerning the specimen placed in the vial. Although onlyone flat portion is shown, the container could be configured without aflat portion, or with two or more flat portions, each adapted to receiveindicia. Alternatively, the indicia could be located on a curved portionof wall 21. The bottom end of flat portion 24 has an arcuate notch 25which acts to keep the container in a proper orientation when handled bythe LBP device, which as noted is designed to cradle the container andmove it through various processing stations. A differently shaped notch(e.g., V-shaped) can be used as long as the notch properly mates withthe LBP device. Other suitable mating structures can be used instead.

Four longitudinal ribs 26 project inwardly from wall 21. The upper ends27 of ribs 26 form rests for the stirrer 40 when it is detached fromcover 30 (see FIG. 10). The top of container 20 has an opening 28 and astandard right-hand helical thread 29 that preferably extends for oneand one half turns and mates with a similar thread on cover 30. Othertypes of cover-to-container coupling may be used, such as a bayonetcoupling, snap-fit arrangement, etc.

Cover 30 comprises a commercially available simple molded plasticthreaded cap 31, and a novel liner 32 retained in the cap. Cap 30preferably is molded of polypropylene, but ABS and EASTAR® DN004, amongothers, are alternative plastic material choices. Cap 31 has a flatsolid top, and an externally knurled depending flange with an internalhelical thread 33 that mates with thread 29 on container 20. Referringto FIG. 4, liner 32 is molded of plastic material, preferablypolyethylene, and has a substantially flat base 34 sized to fit snuglywithin cap 31, behind thread 33, so that the liner is not readilyseparated from the cap. As seen in FIG. 1, liner base 34 serves as agasket-type seal between the cap 31 and the rim of the container wall21.

Liner base 34 has a coupler in the form of an annular projection 35 thatpreferably is slightly conical in shape, preferably forming an angle ofabout 5° to its central axis. In other words, the inner diameter ofannular coupler 35 is greater at its proximal end, where it joins linerbase 34, than at its distal end. Liner base 34 also has a centralannular boss 36 that projects further from base 34 than annular coupler35 so as to interact with stirrer 40, as described below. While the useof a separate liner mated to a standard cap is preferred, the covercould be integrally molded in one piece to include the annular coupler35 and the central annular boss 36. Such a one-piece cover (or even thetwo-piece cover described above) could instead be configured to act as aplug-type seal by projecting into and sealing against the inside of therim of container wall 21.

Referring to FIGS. 1, 3 and 5, stirrer 40 is molded of plastic,preferably polypropylene, and has a circular base or bottom wall 41,sloped at its center, with a central inlet port 42; a central dependingsuction tube 43 with two diametrically opposed suction ports 44 near thebottom of the tube; and a dispersing (mixing) element in the form oflaterally extending vanes 45. The upper portion of the stirrer 40 has acup-shaped particulate matter separation chamber or manifold 46 definedby base 41 and an upstanding annular wall 47. The upper edges of wall 47are beveled, the inner edge 48 preferably being beveled to a greaterdegree to facilitate placement of a filter assembly F in manifold 46, asdescribed below. Possible alternative plastic material for the stirrerinclude ABS and EASTAR® DN004.

Annular wall 47 serves as a coupler for releasably coupling the stirrer40 to cap liner 32, and is therefore dimensioned to fit snugly withinannular coupler 35 (see FIG. 1). Specifically, there is a friction orpress fit between couplers 35 and 47 such that normal handling of theclosed vial, and normal handling of cover 30 when removed from container20 (e.g., to place a biological specimen in the container) will notcause separation of the stirrer from the cover. Coupler 47 isdimensioned relative to coupler 35 so that there is a very slightinitial diametrical interference, preferably about 0.31 mm. Coupler 47is stiffer than coupler 35, so assembly of the stirrer to the coverinvolves slight deformation principally of coupler 35, resulting in africtional force that keeps the stirrer and the cover engaged.Application of an external force to the vial that overcomes thisfrictional retention force will cause stirrer 40 to detach from cover 30and drop by gravity further into container 20 (see FIG. 10).

The external separation force preferably is applied to the centralportion of cover 30 (see the arrow in FIG. 10), which deflects cap 31and liner 32 inwardly. As illustrated in FIG. 1, central boss 36 onliner 32 is dimensioned such that its distal end just contacts or liesvery close to base 41 of the stirrer. Thus, when the central portion ofthe cover is depressed, central boss 36 will deflect further thanannular coupler 35 on liner 32 and push stirrer 40 out of engagementwith coupler 35. Inward deflection of liner 32 also causes coupler 35 tospread outwardly, thereby lessening the retention force and facilitatingdetachment of the stirrer. The separation force applied to cover 30 andrequired to detach the stirrer should be in the range of 5 to 30 lbs.,preferably about 12 lbs.

Once detached from the cover 30, stirrer 40 comes to rest on the upperends 27 of ribs 26. See FIG. 10. The particulate matter separationchamber (manifold) 46 thus is stably supported near the containeropening and easily accessed by the LBP processing heads, which willmanipulate the stirrer so as to process the specimen directly in thecontainer. At least three ribs 26 are required to form a stable supportfor the stirrer, but four are preferred because that number seems topromote more thorough dispersion of the particulate matter in the liquidduring stirring. Should the stirrer inadvertently become detached fromthe cover at the point-of-care site, the physician or an assistantsimply places the stirrer loosely in the vial so that it descends intothe specimen and then screws the cover on as usual. This is notdifficult because the ribs in the vial allow insertion of the stirrer inonly one direction. Once the vial is closed with the specimen inside,the stirrer remains in the vial throughout processing and is sealedtherein when the vial is recapped.

A small percentage of patient specimens, as may be found ingynecological Pap test and other specimen types, contain large clustersof cells, artifacts, and/or cellular or noncellular debris. Some ofthese large objects, if collected and deposited on a slide, can obscurethe visualization of diagnostic cells and, consequently, result in aless accurate interpretation or diagnosis of the slide sample. Sincemost of these features are not of diagnostic relevance, theirelimination from the sample is, in general, desirable. To achieve thisresult, the side suction ports 44 in the stirrer suction tube 43preferably are eliminated (see FIG. 10 a) in favor of close control ofthe interface between the bottom of the suction tube 43 and the smallprojection 23 at the center of bottom wall 22 of the container 20. Thisinterface effectively forms a metering valve whose geometry (orifice) 23a is created when the stirrer 40 rests on the ribs 26 of the container20 (see FIG. 10). Proper sizing of the annular flow orifice 23 aprevents large objects from entering the suction tube 43, while allowingthe passage of smaller objects that may be diagnostically useful. Whilethe orifice 23 a has a thin passage section and a small metering area,clogging is not an issue due to its large diameter. The annular orifice23 a preferably has an outside diameter on the order of 0.105 in. and aninside diameter on the order of 0.071 in., yielding a passage width onthe order of 0.017 in. This orifice size is optimized for gynecologicalspecimens.

Filter System

FIGS. 6 and 8 illustrate one embodiment of a filter assembly F accordingto the present invention. FIGS. 3 and 6 illustrate one embodiment of amanifold 46 (in stirrer 40) according to the present invention. Thefilter system includes the filter assembly F and the manifold 46.

Referring to FIGS. 6 and 8, the filter assembly F comprises a filterhousing or holder 200, a porous frit 202, and a porous membrane filter205. FIG. 8 shows these components more clearly in an exploded view. Theholder 200 can be cup- or container-shaped, having a recess or cavity206 for seating the frit 202 and a chamber 207 between the frit 202 andthe holder 200. The frit 202 and the membrane filter 205 can be made ofthe materials disclosed in the Guirguis patents identified above, namelyU.S. Pat. Nos. 5,301,685 and 5,471,994, the disclosures of which areincorporated herein by reference.

In the present filter assembly F the membrane filter 205, the frit 202,and the holder 200 are assembled together as a unit. The frit 202, whichhas a cylindrical shape, is first seated in the holder 200. Then themembrane filter 205 is permanently affixed, adhered, joined, or fused tothe holder 200. In the illustrated embodiment, the outer perimeter oredge of the membrane filter 205 is fused to the holder 200. In thisregard, the holder 200 has a bevel or chamfer 208 formed around an outercircumferential corner 209. The chamfer 208 provides an angled surfaceto which the membrane filter 205 can be attached using a conventionalbonding technique, such as ultrasonic welding. The holder 200 and themembrane filter 205 should be made of materials that will fuse together.Preferably both are made of polycarbonate, although an ABS holder willwork with a polycarbonate membrane filter. Thermoplastic polyester couldbe used for the holder if the membrane filter is made of the samematerial. The frit 202 preferably is made of polyethylene.

Referring to FIG. 8, the holder 200 preferably is cylindrical andcomprises a substantially cup-shaped body having a bottom wall or base210 and a substantially upright cylindrical sidewall 211 extendingtherefrom and terminating in a rim 211 a. The sidewall 211 has anannular shoulder 212 extending radially inwardly, toward the center. Theshoulder 212 acts as a seat that accurately positions the frit 202. Frit202 preferably is dimensioned so that the frit's outer or front face 213is proud of (extends beyond) the rim 211 a when the peripheral portionof the frit's rear face abuts the shoulder 212.

The inner diameter of the sidewall 211 can be dimensioned tofrictionally engage and hold the frit 202 in place. In this respect, thefrit's outer diameter can substantially correspond to the inner diameterof the sidewall 211 to mechanically, i.e., frictionally, hold the frit202 in place. However, since the membrane filter 205 covers the frit202, the frit need not be frictionally held to the holder. That is, thefrit 202 can be loosely seated in the holder. Frictionally seating thefrit 202 in the holder 200, however, maintains the frit 202 in place sothat attachment of the member filter 205 can be done at a remote site.It also simplifies and reduces the cost of mass production of filterassemblies because the holder 200 and the frit 202 can be joined to makea secure subassembly and stored for later attachment of the membranefilter 205.

After the frit 202 is seated in the holder 200, the membrane filter 205is draped over the frit's outer face 213 and the exposed portion 214 ofthe frit's side wall 215 that extends beyond the holder 200, and isattached to the chamfer 208, as is better seen in FIG. 6. The frit'sexposed outer sidewall portion 214 provides an annular surface areathrough which the specimen liquid can flow to provide a dual flow path,as schematically illustrated in FIG. 7 a.

The filter assemblies F can be coded to denote different pore size andpore density (number of pores per unit cross-sectional area) as may berequired for specific processing protocols. Color coding of filterassemblies is preferred, although any form of machine-detectable codingcan be used, including distinguishing projections, such as smallnipples, for tactile-based sensor recognition. The LBP device isprovided with a sensor that can discriminate between these colors orother codes to ensure proper filter selection. The filter assembliesalso can be provided in paper carriers for easy insertion into the LBPdevice.

Referring back to FIG. 8, the holder's bottom wall 210 has a centralopening 204 through which vacuum can be applied to draw specimen liquidtherethrough. The holder 200 further includes a central projection orprotrusion 216 extending into the holder from the bottom wall 210. Thecentral protrusion 216 is aligned with the opening 204 and positioned inthe chamber 207, which is defined by the frit's inner face 218, theinner face 219 of the bottom wall 210 and the inner side 220 of thesidewall 211. The protrusion 216 is substantially hollow and has aplurality of side openings 221 that distribute vacuum to the chamber 207and provide a substantially symmetrical flow through the chamber. Thespecimen liquid drawn through the membrane filter 205 and the frit 202fills the chamber 207 and exits the chamber 207 through the sideopenings 221 and the central opening 204.

The protrusion 216 has an abutting surface 217 that faces and extendstoward the holder's open face. The abutting surface 217 is configured toabut against the frit's rear face 218. In particular, the abuttingsurface 217 is slightly proud of the annular shoulder 212. That is, theabutting surface 217 lies slightly above or beyond the level of theannular shoulder 212 so that the frit's outer face 213 bows slightlyoutwardly when the frit is installed in the holder. For example, theabutting surface 217 can extend beyond the height of the annularshoulder 212 by about 0.002 inch. The resulting slight bow created bythe protrusion pushing out the central portion of the frit 202 ensuresthat the central part of the membrane filter 205 contacts the slide. Thepressure applied to the slide during imprinting flattens the frit'sfront surface 213, ensuring full contact of the membrane filter 205 withthe slide to more effectively transfer the collected particulates to theslide and minimizing any deposition artifacts. If this slightly bowedconfiguration is desired, the frit 202 preferably is securely seated inthe holder 200, such as by friction as previously explained.

Due to the bowed frit configuration, the membrane filter 205 need not betaut. This simplifies the manufacturing process, reduces cost, andreduces the rejected part rate. Anything short of a major wrinkle canwork effectively. As noted, the frit 202 preferably is slightlydeformable, its compliance allowing it to flex and flatten against aglass slide post aspiration to transfer cells and other objects ofinterest from the filter to the slide. To accomplish this the fritshould have an elasticity that allows it to be crushed flat byapplication of a force of 8 lbs. through a displacement of 0.0016 in.Good frit materials include sintered polyethylene and sinteredpolyester. The frit 202 may be a porous material, with spatially randompores, typically with pore sizes in the range of about 50-micrometer to70-micrometer. A significant attribute of this material is that it is oflow fluidic impedance relative to the material of the thin membranefilter 205 (which typically has pore sizes of about 5-micrometer to8-micrometer). In other words, the pressure drop across the frit 202 ismuch less than the pressure drop across the membrane filter 205. Thus,fluid that passes through the filter flows freely through the frit.Alternatively, instead of having randomly positioned pores, the frit 202may be made of a material or structure that has many parallel channelsof small (e.g., 50-micrometer to 70-micrometer) inner diameters throughwhich aspirated fluid and particulates may flow. Such a parallel-channelarrangement would behave as an inner fluid-pervious medium with anapparent low fluidic impedance. In fact, any material or device with theproper low fluidic impedance and deformability/resiliencecharacteristics may be used in the specimen acquisition station, whetherit has pores or not.

It has been found that flowing the specimen liquid substantially ormostly in an axial direction, i.e., perpendicular to the membranefilter, can accumulate layers or clusters of particulates, asschematically illustrated in FIG. 7 b, particularly if the vacuum isapplied through the membrane filter for a longer period than necessary.This can happen even with the Guirguis dual flow design, which providessome secondary flow components that are radially directed. See, forexample, FIGS. 4 and 12 of Guirguis' U.S. Pat. Nos. 5,471,994 and5,301,685. It seems that the secondary flow generated by thatconfiguration is insufficient to create an effective flushing, orshearing action across the membrane filter. An earlier Guirguis patent,namely U.S. Pat. No. 5,137,031, discloses a funnel- or cone-shapedmanifold. In that arrangement, however, there is no secondary radialoutflow at its periphery. As there is no flow other than directlythrough the filter itself, there is no substantial radial flowcomponent. Accordingly, the specimen liquid only flows substantiallyperpendicularly to the membrane filter.

Referring to FIG. 6, the inner diameter of the upright wall 47 of themanifold 46 at the top of stirrer 40 is dimensioned to be slightlylarger than the outer diameter of the filter assembly F, namely theholder's sidewall 211, so that the manifold 46 can receive and seat thefilter assembly F, with the membrane filter 205 facing down, asillustrated. The filter assembly F can be loosely seated in the manifold46. When the filter assembly F is seated in the manifold 46, the outerperipheral edge of the membrane filter 205 rests on the bottom wall 41.The bottom wall 41 is configured to have a well or recess that forms amanifold chamber M when the filter assembly F is seated in the manifold46. The chamber M is thus bounded by the outer surface of the membranefilter 205 and the upper surface 41S of the bottom wall 41.

The present dual flow arrangement solves the problem of particulatebuild-up or accumulation on the face of the membrane filter. Thisarrangement causes a shearing force or action across the front face ofthe membrane filter that is sufficient to flush the particulates asideand keep them from building up or layering. Built-up or layeredparticulates have a weaker bond to the layer underneath them as theybuild up, because the suction power decreases as the pores of themembrane filter 205 become covered with particulates. A shearing forceis created by imparting a tangential or substantially radial flowcomponent to the specimen liquid across the front face of the membranefilter 205. This flow component is substantially parallel to the frontface of the membrane filter, i.e., it is perpendicular to the built-updirection of the layers, and flushes the particulates radiallyoutwardly, away from the front face of the membrane filter.

To provide a secondary or radial flow path, the manifold 46 isconfigured to provide a small spacing or gap G (see FIG. 6) at theperiphery of the manifold chamber M, between the front face of themembrane filter 205 and the upper surface 41S of the bottom wall 41, toallow flushed particulates to exit the manifold chamber M, away from thefront face of the membrane filter. The gap G must be large enough toprevent the particulates from clogging it. That is, if the gap G is madetoo small for the particulates being filtered, the gap G can getclogged, cutting off the secondary flow. The minimum size of the gapultimately depends on the particulate size, the viscosity of thespecimen liquid, and the temperature of the specimen liquid. It has beendetermined that the gap G should be at least 0.004 in. to preventclogging by cellular particulates.

Referring to FIGS. 3 and 6, to create the gap G, which forms an outflownozzle, the bottom wall 41 of manifold 46 includes a plurality of spacedstandoffs or raised ribs 48 a around the periphery of the manifold 46.The spaces 49 between the ribs 48 a provide a passage for specimenliquid to exit the chamber M. In the illustrated preferred embodiment,the manifold 46 has an inner diameter of 23.4 mm, and has thirty-sixribs 48 a, evenly spaced at 10°. The ribs are 0.150 mm high andarcuately blend into the surrounding shoulder with a radius R of 0.63mm, as illustrated. Of course, the present invention contemplates otherconfigurations of spaced ribs or standoffs, which are intended toprecisely space the filter assembly from the bottom wall 41 so that aprecise outflow area is created. Depending on the number and thicknessof ribs or standoffs, the total outflow area can be reduced as much as50% as compared to the inlet area.

It has been observed in the Guirguis type filter arrangement referred toabove that specimen liquid traveling radially outwardly loses velocity.The present dual flow filter system compensates for the velocityslowdown by providing a shallow, substantially conical surface acrosswhich the specimen liquid flows. This surface forms a substantiallyconical distribution manifold chamber M confronting the membrane filter205. The chamber M according to the present invention has an annularradial outlet O, through spaces 49, having an area that is about equalto or smaller than the maximum area of the central inlet I. Referring toFIG. 9, the “face” area of the radially directed annular flow passage iscylindrical and is defined (bounded) at any given radius R₁, R_(x),R_(y), . . . , R₂ by the front surface of the membrane filter 205 andthe conical surface 41S of the manifold. As the specimen liquid travelsoutwardly, the radius increases while the manifold height decreases. Themanifold chamber M can be configured so that the height H₁, H_(x),H_(y), . . . , H₂ decreases at a rate which maintains the face area ofthe annular passage substantially uniform from the inlet I to the outerperimeter outlet O of the manifold, yielding a substantially linearradial flow velocity across the face of the membrane filter 205.

In this regard, still referring to FIG. 9, the maximum theoreticalradial flow area of a round manifold inlet I can be defined as thecircumference (2πR₁) multiplied by the height of the manifold chamberH₁. In this instance, 2πR₁H₁ defines the total circumferential area ofthe manifold inlet I. The maximum circumferential flow area of a roundmanifold outlet O can be defined as 2πR₂H₂. If the outlet flow area isto equal the inlet flow area, then the inlet and outlet areas can beexpressed as:2πR₁H₁=2πR₂H₂R₁H₁=R₂H₂Using this expression, the heights, e.g., H_(x), H_(y), can be definedat their given radii, e.g., R_(x), R_(y) from the inlet I to the outletO. If the heights H₁, . . . , H_(x), . . . , H_(y), . . . H₂ from theinlet to the outlet are plotted, the resulting surface 41S would becurved, not linear. However, it has been observed that a significantlycurved lower manifold surface does not work as effectively as a linearsurface 41S. Accordingly, the present preferred embodiment contemplatesa linear or substantially or nearly linear surface 41S (which can beslightly curved) extending from the inlet to the outlet. Also, there isa minimum height H₂ of about 0.006 inch clearance for the specimenliquid to effectively flow. Based on this requirement, the minimum R₁can be defined as 0.006R₂/H₁ inches. With this configuration, as thespecimen liquid is drawn through the filter, the specimen liquidtraverses the front face of the membrane filter 205 in a direction thatis substantially parallel to or approaching nearly parallel to the frontface of the membrane filter, creating the desired shearing action.

Empirical study has revealed that for a linear conical surface 41S, thearea of the outlet O preferably should be less than or equal to themaximum area of the inlet I. That is, R₁H₁ R₂H₂. For example, theexemplary manifold can have the following dimensions (all units here inmm): R₁=1.24, H₁=1.32, R₂=10.00, H₂=G=0.15. The maximum inlet area wouldthus be 3.277 πmm² and the outlet area 3.00 πmm², which is slightly lessthan the maximum inlet area, but greater than the average inlet area,which can be defined as 50% of the maximum inlet area (1.64 πmm²). Thus,the outlet area can fall between the maximum inlet area and the averageinlet area. Another example can have the following dimensions (all unitshere in inches): R₁=0.040, H₁=0.060, R₂=0.400, H₂=0.006. The maximuminlet area would thus be 0.0048 πin², which is equal to the outlet area.

In summary, the manifold chamber M that confronts the substantially flatmembrane filter should have a shallow, funnel-shaped configuration and aperipheral outlet so as to create a substantial radial flow across theouter surface of the membrane filter. The radial flow creates a shearingaction that washes or flushes away any particulates that are relativelyweakly attached so as to leave a very thin layer of particulates—amonolayer—on the surface of the membrane filter.

LBP Device and Method

FIGS. 11-57 illustrate a preferred embodiment of an LBP device accordingto the present invention. The LBP device is an automated machine forpreparing slides for viewing, imaging or optical analysis. The LBPdevice can use the above-described dual flow filtering system (FIGS. 6,7 a, 9) to collect monolayers or thin layers of cells and transfer themonto slides.

Referring to FIG. 11, the illustrated embodiment of the LBP device canbe compartmentalized into at least six discrete processing stations:data acquisition station (bar code reader) 230; uncapping station 400;primary stirring station 500; filter placement station 600; specimenacquisition station 700; and re-capping station 800. These six stationsare structured for parallel processing, meaning that all these stationscan operate simultaneously and independently of the other. The LBPdevice also includes a separate data reading station, a slidepresentation station, a slide handling station, and a cassette handlingstation, all of which can be incorporated as an integrated system 900.The LBP device further includes a transport mechanism 240 for moving thespecimen containers to the various operating stations. It can furtherincorporate an auto loading mechanism 300 that automatically loads andunloads specimen vials onto and from the transport mechanism. Allstations are computer-controlled. FIG. 11 a shows the operating sequenceof the LBP device. This is the top-level table from which the operatingsoftware is structured.

FIG. 12 shows the basic structural elements of the LBP device, namely aframe 260 preferably made of extruded aluminum, preferably on casters(not shown) for mobility, and a machined aluminum base plate 262supported by the frame and on which the main operating mechanisms aremounted. Beneath the base plate is a compressor 264 for supplyingcompressed air for powering some of the components; a vacuum pump (notshown) which provides a vacuum source for various components; stainlesssteel shelves for holding the vial trays used in the auto loadingmechanism 300; and electrical components, including power supplies andcontrollers, and miscellaneous equipment. A compressor would not berequired if electrically-powered actuators were used instead ofair-powered actuators. A user interface, e.g. a touch-sensitive LCDdisplay (not shown), is mounted to the left of the transport mechanism240 and gives the technician control over machine operation beyond thenormal automated processing protocols. See FIG. 25, which shows examplesof a log-in screen (top) and a navigation screen (bottom) as they mightappear on the user interface. Of course, other screens would bepresented to the user as he/she interacts with the user interface.

An “economy” version of the LBP device can take the form of acounter-top model for processing a more limited number of specimens at atime. In such a model certain components can be eliminated, such asframe 260 and auto loading mechanism 300, while other components can bescaled back, such as the capacity of filter placement station 600.External sources of vacuum and compressed air could be used to powersuch a device, while other components (power supplies, controllers,etc.) could be repositioned to one or more modules adjacent to or on amodified machine base plate. Various ways of implementing thesemodifications will be readily apparent to those skilled in the art.

Transport Mechanism

Referring to FIG. 11, the transport mechanism 240 comprises an endlesslink-belt conveyor 242 driven by a stepper motor (not shown) aroundprecision sprockets 242, 244. The conveyor has a plurality ofreceptacles or carriers 246, linked by pins 248, for receiving acorresponding number of specimen vials. The illustrated embodiment inFIG. 11 has 30 receptacles, numbered 1 through 30. Depending on thesample vial size and the length of the conveyor, the LBP device can usefewer than or greater than 30 receptacles, as desired or feasible,sufficiently long to permit all processing to be completed in a singleline.

The receptacles 246 of the link-belt conveyor are guided between thesprockets by pairs of guide rails 250 forming tracks, and has aconventional position correction system (not shown) to accuratelyposition the receptacles. The LBP device can track the position of eachreceptacle and step-drive or index them in a conventional manner. Forinstance, the LBP device can include linear position sensors, such asoptical sensors or a photo-interrupter on each link, that can feed theposition to a controller for registering carrier position and preciselyindexing each carrier at each of the processing stations along theprocessing path. The manner of driving the conveyor for precisealignment and positioning is conventional and thus will not be describedfurther.

The guide rails 250 that form tracks in Z and Y axes engage slotsmachined in the sides of the receptacles. See, for example, FIGS. 29,33, 37 and 43. The mechanical tracks and drive sprockets can beconstructed of a self-lubricating plastic for operation without the needto add an external lubricant. The receptacles 246 each can have a window247 (see FIG. 12) for allowing access to laser or optical scanning ofthe bar code on the specimen containers. The conveyor can be hard-coatedaluminum, ®-impregnated with PTFE7 for easy cleaning. The link pins 248can be precision ground and hardened. The link pins can be axially fixedin location in the non-rotating link bore. Rotating link bores can befitted with a suitable bearing material capable of operation withoutadditional lubricant. For operator safety, the conveyor operation can beinterlocked with the cover of the machine (not shown).

The receptacles 246 are also configured so that they receive or seat thespecimen vials in a particular orientation. That is, the specimen vialsand the receptacles are complementarily configured or keyed so that thevials can only be seated in the receptacles in a particular orientation.For example, the vials can be “D” shaped, namely having a flat side (seeFIGS. 2 a, 2 b), and the receptacles can be “D” shaped so that the flatsides align with each other. In this way the vials do not rotaterelative to the receptacles, while allowing unrestricted verticalmovement relative to the receptacles. In addition to the D shape, eachvial can have a bottom notch 25 (see FIG. 2 a), and the receptacles canhave a mating peg or stud (not shown) that keys into the notch 25. Whilethe illustrated notch and peg are arcuate, they can take on other matingshapes (e.g., V-shaped).

Vial Loading/Unloading Mechanism

FIGS. 12, 13 and 14 show the automated vial loading and unloadingmechanism 300. A pivoted pick-and-place arm 304 is mounted on anelevator carriage 306 driven by a vertical (Y-axis) lead screw motor 308atop a vertical standard 310. Arm 304 has a conventional electrically-or pneumatically-operated jaw-type gripper 312 adapted to grasp and movespecimen vials 10 in three degrees of freedom. Arm motion in horizontalplanes is afforded by lateral lead screw motor 314, which is pivotallymounted in a clevis-type bracket 316 to elevator carriage 306. Insteadof a jaw-type gripper as shown, the pick-and-place arm can be equippedwith a conventional pneumatically operated suction-head type gripper asshown in FIG. 15. Such a gripper has a silicone rubber bellows 318 whichseals against the cover 30 of a vial when placed against the cover andsubject to suction through a suction line 320. Whether mechanical orpneumatic, actuation of the gripper is accomplished through theprogrammed operation of the machine as is understood by those skilled inthe art.

Referring to FIGS. 17-20, specimen vials 10 are stored in specialinjection molded plastic vial trays 330 that slide into the machine onshelves 320 (see FIG. 12). To avoid confusion, it should be pointed outthat FIGS. 13-15 show a different form of tray (made of stamped steel),but the operation of the mechanism that rotates the trays, regardless oftheir construction, is the same. The plastic vial trays 330 are thepreferred form, and are preferably made of polypropylene. The term“tray” as used herein is not limited to the embodiments shown, andshould be construed to cover any type of carrier, rimmed or rimless,that can support and move a generally planar array of discrete articlesgenerally in the manner described herein.

Each tray 330 has forty-one circular recesses 332 sized and configuredto receive specimen vials 10 only in one orientation. The upper edge ofeach recess 332 preferably has a beveled edge 333, which facilitatessmooth insertion of vials. The recesses are arranged in a close-packarray of four concentric rows, preferably as follows. The outermost rowhas sixteen recesses; the next row in has eight recesses; the third rowin has nine recesses; and the innermost row has eight recesses. Thereceptacles of adjacent rows are offset for closer spacing. Thereceptacles of the second row are radially aligned with the receptaclesof the fourth (innermost) row. The receptacles of the outermost row arespaced at 18° on center. The receptacles of each of the other rows arespaced at 36° on center. Of course, other receptacle arrays could beused as long as they permit access of all vials by the pick-and-placearm 304. Each receptacle has a unique and addressable location, so thatany vial can be accessed at will and in any sequence.

As noted above, orientation of specimen vials during the processing iscritical, so the proper orientation of the stored vials in these traysensures that the pick-and-place arm 304 will properly position each vialin a conveyor receptacle 246. Accordingly, each recess 332 has at itsbottom (see FIG. 19) a fixed indexing peg 334 that is sized to fit intonotch 25 in the vial. The pegs 334 are installed, e.g., by adhesive, ingrooves 335 that are molded into the tray adjacent the bottoms of therecesses 332. Some of the pegs have been omitted from FIG. 19 forillustrative purposes.

The pegs 334 are arranged at specific angles with respect to the medianplane of the tray 330 such that each vial removed from the tray isdelivered to a transport receptacle with its notch aligned with themating peg in that receptacle, and vice versa. Each of these angles isdictated by the rotational position of the tray 330 when a vial in aspecific recess 332 is to be accessed by the pick-and-place arm 304, andthe angular rotation of the pick-and-place arm from the point of vialpick-up to the point of vial placement in the conveyor receptacle 246.The determination of these angles is considered to be within theabilities of one of ordinary skill in the art.

The tray 330 also has three upstanding guide posts 336, each with aspring-loaded ball 338 at its tip, which cooperate with guides (notshown) above each shelf 302 and serve to guide the tray into the machineas it is inserted and ensure its proper orientation. The guide posts 336also serve as stacking posts when the trays are stacked for storage (seeFIG. 20), the balls 338 engaging dimples 339 (see FIG. 19) in the bottomof the superior tray.

The tray 330 also has a large flared notch 340 which is oriented towardthe machine when the tray is inserted on a shelf 302. The innermostportion of the notch 340 has opposed keyways 342 which are adapted forengagement by floating keys, as described below. The keyways preferablyare formed in a milled brass hub insert 343 that is recessed flush withthe top of the tray and secured thereto by screws.

Referring to FIGS. 14, 15 and 15 a, a rotary outer spindle 350 isjournaled at its top and its bottom in bearings 352, 354, respectively.Outer spindle 350 engages and rotates only one tray at a time so thatthe pick-and-place arm 304 can access vials therefrom by movingdownwardly through an opening 266 in base plate 262 and past any idletrays via their homed notches 340. FIG. 14 shows the home positions ofthe trays in dashed lines, with their notches 340 aligned and embracingouter spindle 350. Spindle 350 is rotated in a precision manner from thebottom by a computer-controlled rotation stepper motor 356 and a timingbelt 358 engaging timing gears 360, 362. A downwardly facing opticalrotary position sensor 363 located over the aligned tray notches detectswhen and how far a tray is rotated from its home position and providescontrol feedback for rotation of stepper motor 356.

Within outer spindle 350 is an inner spindle 364 carrying eight pairs ofopposed keys 365, one pair for each tray. The keys 365 project fromouter spindle 350 through opposed slots 366 in the outer spindle (seeFIG. 15 a, which is a sectional view through the spindles and the centerportions of the bottom two trays). The inner spindle 364 is movedvertically within the outer spindle 350 by an internal lead screw 372.Lead screw 372 is rotated by lead screw stepper motor 374 through atiming belt 376 and timing gears 378, 380. A key “home” sensor 382 (seeFIG. 15) is located at the top of inner spindle 364 to provide areference point, i.e., when the machine is turned on, it will “home” theinner spindle to the key home sensor 382 and then reference itsmovements from there.

The even vertical spacing of the pairs of keys can be seen in FIG. 15.This spacing, or pitch, differs from the pitch of the keyways 342 in afull complement of installed trays 330. Accordingly, which keyways areengaged by the keys depends on the vertical position of inner spindle,and only one pair of keyways (tray) can be engaged at any time. Theenlarged view of FIG. 15 a shows that the keyways 342 of bottom tray330-1 are engaged by keys 365, while the keyways of the tray above it,330-2, are not engaged by any keys. Movement of inner spindle 364 byone-eighth the pitch difference disengages one tray and engages theimmediately adjacent tray. The operation of the loading and unloadingmechanism is unaffected by the absence of one or more trays from thetray slots, which are defined by shelves 302.

When a selected tray is to be accessed by the pick-and-place arm 304 (asdetermined by the computer controller), the lead screw motor 374 movesthe inner spindle the appropriate distance so that the appropriate keysengage the keyways of the selected tray. The rotation motor 356 thenrotates the keyed tray to the proper angular position so the arm 304 canaccess a particular recess 332. The superposed arrangement of the trays,the way in which a selected tray is accessed by the gripper 312 throughthe flared notches 340 of superior trays, and the close-pack spacing ofthe recesses 332 in each tray make for an extremely compact, highcapacity and efficient vial handling system that is readily incorporatedinto the compact base of the LBP device.

In the embodiment shown, the LBP device can accommodate up to eighttrays holding forty-one specimen vials each. One of the forty-onerecesses can be reserved for a cleaning vial, which would contain acleaning solution and be run through the LBP device to clean the variousparts of the device that normally come into contact with specimen fluid.Alternatively, the forty-first vial could contain a typical controlspecimen for calibration purposes. Thus the LBP device can accommodateup to at least 320 vials containing specimens to be processed. Thedevice is therefore capable of operating continuously unattended for along duration—at least eight hours—so that specimen processing can becarried out even when laboratory personnel are not normally present,such as at night.

When the trays 330 are bar-coded or otherwise labeled withmachine-readable identifying data, they can be used in an automatedstorage device that can access a particular tray on command. Thetray-identifying data can be input into the integrated data managementsystem so that the location of any specimen vial in tray storage can bereadily ascertained.

A cost reduction in tray-based storage of specimen vials can be achievedby using a liner-type system in conjunction with trays 330. For example,vials can be supported and stored in thin sheet-like liners (not shown)that conform to trays 330 and slip readily into recesses 332. The linersare stiff enough to be self-supporting when fully loaded, can bestacked, and can be housed in wheeled carts for ease of mobility.

Data Accessioning and Specimen Management

It is, of course, important to keep track of each specimen vial and thespecimen slides produced from each vial. Accordingly, the LBP devicetypically communicates with the integrated data management system (DMS)104 through an accessioning station 102 or other computer. FIG. 21schematically illustrates specimen vial handling and the flow of datathat is integrated into to operation of the LBP device. Thecommunication link between the LBP device and the DMS can be made viaethernet or other protocol using a direct peer-to-peer connection, orthrough a server-based network.

The specimen processing operation begins with collecting or transferringdata from the labeled specimen vial, e.g. via a bar code reader on adata entry terminal or accessioning station, to the DMS via either adirect connection or over a network. Specimen tracking data can include,for example, the patient's name, test identification (ID) number,patient data, and any special processing instructions. For example, thebar-coded specimen vial can be linked to the patient informationinitially by a paper requisition form and subsequently by an assigned,unique numerical ID in the database. In a preferred embodiment, thepatient and test information including the vial bar code can be enteredinto the networked DMS database at the point-of-care site (e.g.,physician's office), thereby eliminating entirely the need for a paperrequisition form. U.S. Pat. No. 5,963,368 (incorporated herein byreference), which is assigned to AccuMed International, Inc. (nowMolecular Diagnostics, Inc., or MDI) discloses a similar concept asapplied to a computer-controlled instrument for analyzing biologicalspecimens (a microscope) and storing data from each analysis. The '368patent is exclusively licensed to MonoGen, Inc. (the owner of thisapplication) in the field of liquid-based cytology in combination withor for use with non-fluorescence based image analysis devices,processes, systems and/or instruments. MonoGen's commercially availablepathology work station and data management system implement the conceptdisclosed in the '368 patent.

Each specimen vial includes an identification (ID) symbol or label(e.g., bar code) and/or a stored information label or symbol such as ahologram or a memory chip or device. The present embodiment contemplatesreading an ID label using an optical reader, such as a bar code reader,which provides the information to a DMS for sharing information betweendifferent work stations or instruments at the same or differentlocations, such as laboratories, doctors' offices, hospitals, or otherpatient care providers. FIG. 21 a depicts an overall laboratory systemwherein the DMS is expanded to link specimen/patient data through aserver to a variety of specimen processing devices and/or computerizedwork stations for fully integrated specimen management.

A separate bar code reader 230 (see FIG. 11) is mounted on the LBPmachine itself, and scans all specimen vials prior to processing througha slit in each transport receptacle 246. Each of the transportreceptacles 246 is tracked using this symbol or code, such as a bar codethat can be read with a conventional optical reading device. The barcode readers used in the LBP device can be any commercially availabletype, such as Keyence BL-600, with a minimum BCR target code capabilityof Interleaved 2 of 5, Code 128c, or EAN-128. The bar code readerspreferably are sealed in liquid-tight enclosures for operatorprotection. After reading, specimen vial/transport receptacle ID dataare transmitted to the DMS of the host database or work station. Thehost database or local work station can then transmit back to the LBPdevice the specific processing protocol to be performed on thatindividual specimen.

Some of the most important functions of the data management system (DMS)include:

-   -   Obtaining data on the patient and the specimen during        accessioning, and making this available to each instrument as        required to set processing parameters and to provide medical        data to the slide reviewer;    -   Maintaining chain of custody of specimens and slides to ensure        data integrity;    -   Compiling data and printing required forms for regulatory,        compliance, and laboratory management reports;    -   Generating medical reports and ensuring integrity using        safeguarded digital electronic signatures;    -   Managing billing for instruments on “per use” charges;    -   Storing optimal processing protocols for each process and        supplying to the instrument in accordance with the specimen type        and/or user requirements; and    -   Facilitating remote diagnostics and repair, and providing user        manuals and troubleshooting guides.        FIG. 21 b shows an example of a relational database table that        can be used to accomplish these tasks.

The DMS can provide paper-free data flow among the different stages ofthe cytology process, saving a significant amount of personnel time andcost, reducing transcription errors, improving accuracy, and eliminatingthe space required to store paper records. By automating and managingdata acquisition, storage and retrieval, each operation becomes moreefficient, significantly reducing the turn-around time for specimens.Specimen quality is enhanced by automated calibration and cross-checkingroutines that identify potential problems early. Flexible foreignlanguage support for worldwide sales assists laboratories inmulticultural environments.

The DMS provides a common user interface that provides detailedinformation on the operation of each connected laboratory device andwork station, and together with online user manuals and training aidseases use and minimizes training. The DMS handles the exchange of allrelevant patient and specimen data with the users' own LIS (or otherdata management systems) through a provided software interface.Moreover, remote instrument diagnostic capabilities ensure maximuminterruption-free operation. The reduction in paperwork, readycross-compatibility with other instruments and existing computernetworks, and integration with the central hospital or laboratoryinformation system provides significant user benefits.

In typical operation, the laboratory: (1) receives a requisition fromthe healthcare provider along with the pre-bar-coded specimen vial, (2)assigns a unique ID number (accession number) to the specimen, and (3)based on information on the requisition, enters a specific LBP test IDto specify the process to be used. FIG. 23 shows an example of theaccessioning (data entry) screen that is presented to the technician,into which the vial bar code, accession number and LBP process code areentered. When the specimen vial is loaded into the LBP device forprocessing, the LBP device automatically reads the bar code on thespecimen vial and transmits the bar code number (106) to the DMS, whichsends back the processing parameters for the selected test, and thenumber of slides to be produced. The LBP device returns anacknowledgment (108) and processes the specimen, making one or moreslides as instructed via the DMS. Immediately before the LBP deviceimprints a specimen slide with material filtered from a specimen vial,the LBP device reads the bar code from the pre-bar-coded slide that isto receive the specimen sample. The LBP device sends each slide bar code(110) and its associated vial bar code to the DMS which updates thepatient database with the slide bar code number, cross-references it tothe correct vial number, and signals (112) the LBP device to proceed.The LBP device then imprints a cytological sample from the specimen ontoone or more slides and readies the onboard data log for the nextspecimen to be processed. FIG. 24 shows an example of a DMS menu screenshowing data items that are now linked in the DMS database, includingthe vial number, slide number(s) and patient data. The DMS can produce aprintable report listing slide ID numbers and associated vial IDnumbers, patient data and processing protocols.

At a minimum the protocol variables include specimen mixing parameters(stirring speed and time) and filter selection. Typically, primarystirring speed can be varied from 500 rpm to 3,000 rpm selectable in 50rpm steps. The stirring interval can be varied from 5 to 120 seconds,selectable in 5 second increments. Choice of filter type is based onaverage pore size diameter: either 5 micron (red housing), e.g. fornon-gynecological specimens, such as sputum specimens, or 8 micron(white housing), e.g. for gynecological specimens, depending on the testprotocol selected.

The LBP device is capable of processing mixed sample-runs (i.e., runsthat may include vials containing various types of specimens)interchangeably and without the need for batch processing of same-typespecimens. Specimen processing can include at least 100 differentprocessing protocols resident within the DMS and accessible to users.Predefined procedure codes (test ID's) such as the following can be usedto simplify operator input and specify which processing protocol isused:

-   -   1 breast cyst, L    -   2 breast cyst, R    -   3 bronchial brushing    -   4 bronchial washing    -   5 bronchoalveolar lavage    -   6 cerebrospinal fluid    -   7 colonic brushing/wash    -   8 esophageal brushing/wash    -   9 gastric brushing/wash    -   10 gingival (buccal) scrape    -   11 gyn PAP test    -   12 intestinal brushing/wash    -   13 nipple discharge, L    -   14 nipple discharge, R    -   15 ovarian cyst, L    -   16 ovarian cyst, R    -   17 pericardial effusion    -   18 peritoneal effusion    -   19 pleural effusion    -   20 rectal brushing/wash    -   21 sputum, induced    -   22 sputum, spontaneous    -   23 urine, catheterized    -   24 urine, voided

Each specimen is processed with a new filter to prevent the possibilityof cross contamination. In the present embodiment, either of two or moredifferent filter types can be specified for versatility in testselection (the device's eight filter tubes allow for up to eightdifferent filter types). Processing parameters for each type of specimenpreparation can be determined remotely and in advance, and communicatedto the processing device using a bi-directional communication linkutilizing the specimen vial bar code as the key identifier. The LBPdevice can utilize default (pre-loaded into the DMS) process protocolsas well as laboratory-generated process protocols that users can add tothe DMS.

An overfilled-vial sensor (not shown) can be positioned at or justdownstream of the bar code reader 230 to detect whether an excessiveamount of fluid is present in each translucent vial. Opening andprocessing an overfilled vial can result in hazardous spillage orejection of biological fluid. Accordingly, if an overfilled vial isdetected, the DMS will be so notified and the complete LBP processingprotocol for that vial will be canceled, allowing the overfilled vial toproceed through the processing path unopened. Alternatively, anoverfilled condition can be sensed at the conveyor holder 246 into whichvials are loaded by the vial loading mechanism 300. If an overfilledvial is detected there, the DMS will be so notified and the loadingmechanism will be instructed immediately to return the overfilled vialto its tray 330.

A similar approach can be used to deal with other anomalies detected aseach vial is loaded into the conveyor. For example, a sensor (not shown)can be used to detect an unreadable bar code on the vial, or detect whena vial is improperly position in the holder 246. When any such conditionis detected, the DMS will be so notified and the loading mechanism willbe instructed immediately to return the overfilled vial to its tray 330.

FIG. 22 is a block diagram showing the components of a general purposecomputer system or work station 270, which can be used to run the DMS.The computer system 270 typically includes a central processing unit(CPU) 272 and a system memory 274. The system memory 274 typicallycontains an operating system 276, a BIOS driver 278, and applicationprograms 271, such as a DMS. In addition, the computer system 270 caninclude input devices 273, such as mouse, keyboard, microphone,joystick, optical or bar code reader, etc., and output devices, such asa printer 275P, and a display monitor 275M.

The computer system or work station can be connected to an electronicnetwork 280, such as a computer network. The computer network 280 can bea public network, such as the Internet or Metropolitan Area Network(MAN), or other private network, such as a corporate Local Area Network(LAN) or Wide Area Network (WAN), or a virtual private network. In thisrespect, the computer system 270 can include a communications interface277, such as ethernet, USB, or Firewire, which can be used tocommunicate with the electronic network 280. Other computer systems 279,such as a remote host database, other types of work stations includingautomated analyzers, and computers or databases (e.g., LIS) of ahospital, laboratory, or other medical establishment, can also be linkedto the electronic network 280. Other LBP devices, as well as other typesof specimen processing instruments (e.g., automated slide stainers andcoverslippers) 279 a can also be connected to each other and the DMS viathe network.

One skilled in the art would recognize that the above-described systemincludes typical components of a general purpose computer systemconnected to an electronic network. Many other similar configurationscan be used to control the LBP device and its processes. Further, itshould be recognized that the computer system and network disclosedherein can be programmed and configured by one skilled in the art toimplement the methods, system, and software discussed herein, as well asprovide requisite computer data and electronic signals to implement thepresent invention.

In addition, one skilled in the art would recognize that the “computer”implemented invention described further herein may include componentsthat are not computers per se, but include devices such as Internetappliances and Programmable Logic Controllers (PLCs) that may be used toprovide one or more of the functionalities discussed herein.Furthermore, while “electronic” networks are generically used to referto the communications network connecting the processing sites of thepresent invention, one skilled in the art would recognize that suchnetworks could be implemented using optical or other equivalenttechnologies. One skilled in the art would recognize that other systemconfigurations and data structures can be provided to implement thefunctionality of the present invention. All such configurations and datastructures are considered to be within the scope of the presentinvention. In this context, it is also to be understood that the presentinvention may utilize known security and information processing measuresfor transmission of electronic data across networks. Therefore,encryption, authentication, verification, compression and other securityand information processing measures for transmission of electronic dataacross both public and private networks are provided, where necessary,using techniques that are well known to those skilled in the art.

Uncapping Station

One of the advantages of the present vial-based LBP device and system isthat it minimizes operator exposure to the specimens, which can containpotential biohazards. Referring to FIGS. 26-31, the LBP device has anuncapping mechanism 400 that first automatically separates the stirrer40 in the vial from cover 30, and then removes and discards thecover—all without intervention by an operator. See FIG. 26, which showsthe stirrer resting on vial ribs 26 after the cover 30 is removed.

A closed specimen vial 10 which has arrived at the uncapping station inits transport receptacle 246 is met by an uncapping head 402 which islowered onto the cover 30 of the specimen vial. See FIGS. 27 and 28.Uncapping head 402 has four tapered legs 404 that form a taperedgripping cavity having chisel-like inner edges 406 spaced and sized toprogressively tighten onto cover 30 as head 402 is lowered. Once thecover is tightly engaged by the legs, a central spindle or plunger 408is lowered into contact with the center of cover 30 and applies adownward force to the cover to cause the stirrer 40 to detach from thecover 30, as described above, and drop down in the vial onto ribs 26.Then the plunger is retracted and the uncapping head 402 is rotatedcounterclockwise (FIG. 28) to unscrew cover 30 and remove it fromcontainer 20. Thereafter the uncapping head with the removed cover inits grip moves laterally to the position shown in dashed lines 410 inFIGS. 29 and 11, and plunger 408 is again lowered, this time to ejectcover 30, which falls into a waste chute or bin (not shown) beneath theuncapping head. Alternatively, a movable waste chute can be broughtbeneath the uncapping head to catch the ejected cover, so that lateralmovement of the uncapping head is not required. Covers are not reused toeliminate the possibility of cross-contamination.

The plunger 408 is driven by a pneumatic cylinder 412, mounted on anL-bracket 415 at the top of the uncapping head, that can apply a forceon the cover of up to about 30 lbs. A coil spring 413 returns theplunger to its retracted position when cylinder 412 is deactivated. Thehead 402 is capable of applying an uncapping torque through the grippinglegs of up to about 10 lb-ft, which is sufficient to loosen the cover.The gripping legs can be of the self-energizing type so that precisealignment with the cover or small variations in cover geometry do notfrustrate their grip.

The uncapping mechanism has a mounting frame 414 supported on blocks 416that slide laterally of the processing path on rails 418. A Y-axisstepper motor 420 and lead screw 422 effect lateral motion. Theuncapping head 402 is rotatably mounted in a bearing block 424. Bearingblock 424 is secured to a C-frame 426 that is vertically slidable onmounting frame 414. Vertical movement of C-frame 426 and, hence,uncapping head 402 is effected by Z-axis stepper motor 428 and leadscrew 430. Lead screw 430 can be vertically compliant to accommodateupward movement of the cover 30 as it is unscrewed. However, it ispreferred that stepper motor 428 be actuated during the uncappingsequence so that head 402 rises at about the same rate as, but no fasterthan, the unthreading cover. Uncapping head 402 is rotatably driven byuncapper motor 432 through a gear reduction unit 433, a timing belt 434and timing pulleys 436, 438.

The uncapping head described above would also work with vials having aconventional “press and turn” bayonet-type coupling between thecontainer and the cover. The downward force of the plunger 408 would besufficient to release the internal anti-turn lock of the coupling,allowing the gripper to rotate and remove the cover. Vials having coversthat do not require rotation for removal, e.g., a snap-on cover, wouldrequire a differently designed uncapping head, tailored to the type ofcover connection involved.

Alternatives to the above-described plunger 408 can be employed at orupstream of the uncapping station for applying the required externalforce to the covered vial to effect separation of the stirrer from thecover. For example, a cam, lever arm or other movable mechanical elementcan contact and press down on the cover. Alternatively, an abrupt upwardexternal force can be applied to the vial to yield an acceleration forcethat overcomes the frictional retention force between couplers 35 and47, effectively pulling the stirrer out of engagement with the cover.This can be done by, e.g., moving the closed vial rapidly downwardly torap the bottom of the container 20 against a rather hard surface, e.g.,by mechanically and/or pneumatically thrusting the closed vial into thetransport carrier 246 that will hold the vial during the subsequentprocessing steps, or by dropping the vial down a chute into the carriera sufficient distance to dislodge the stirrer. Another way to exert anabrupt upward external force on the vial is to strike the bottom of thecontainer 20 with a striking member. This can be accomplished by, e.g.,cradling the container 20 and momentarily thrusting a striker againstthe bottom of the container, e.g. through a bottom opening in the vialcarrier 246, by pneumatic and/or mechanical means. The design of theseand other variants of suitable automated mechanisms for accomplishingthese tasks is within the grasp of those skilled in the mechanical arts.

Preprocessing (Primary Stirring) Station

After uncapping is completed, the transport mechanism indexes thespecimen container to a station where preprocessing occurs. Thepreprocessing station is the location at which preprocessing operations,such as specimen dispersal within its container, are performed prior tothe container and its specimen moving to the specimen acquisitionstation. The preprocessing station typically performs a dispersaloperation. In the preferred embodiment, the dispersal operation isperformed by a mechanical mixer, which rotates at a fixed speed and fora fixed duration within the specimen container. In this example, themixer serves to disperse large particulates and microscopicparticulates, such as human cells, within the liquid-based specimen byhomogenizing the specimen. Alternatively, the specimen may containsubcellular sized objects such as molecules in crystalline or otherconformational forms. In that case, a chemical agent may be introducedto the specimen at the preprocessing station to, for example, dissolvecertain crystalline structures and allow the molecules to be dispersedthroughout the liquid-based specimen through chemical diffusionprocesses without the need for mechanical agitation. In this example,the chemical preprocessing station introduces its dispersing agentthrough the preprocessing head.

In the illustrated preferred embodiment preprocessing occurs at theprimary stirring station 500, which uses a specified or instructedstirring protocol to stir the specimen, if needed, using the stirrer 40in the container, at a specified speed (rpm) for a specified duration.The stirring protocol chiefly depends on the specimen, as describedabove, and is normally intended to disaggregate any mucous material anddisperse it and/or other particulate material in the specimen liquid.

Referring to FIGS. 32-35, the primary stirring station 500 has astirring head 502 in the form of an expanding steel collet. The colletis formed at the lower end of a shaft 503 which splits into six flexiblefingers 504 defined by six equally spaced slits 506. Shaft 503 isrotatable in a bearing block 508 secured to a C-frame 510 that isvertically slidable on a mounting frame 512. Vertical movement ofC-frame 510 and, hence, stirring head 502 is effected by a Z-axisstepper motor 514 and a lead screw 516. Stirring head 502 is rotatablydriven by a stirring motor 518 through a timing belt 520 and timingpulleys 522, 524.

The inner surfaces of the collet fingers 504 taper uniformly inwardlytoward the lower end of the collet. A central plunger 526, movablevertically by a pneumatic cylinder 528 atop a bracket 530, expands thefingers 504 outwardly when it descends and encounters the narrowingpassage defined by the tapering fingers. Thus the diameter of the lowerend of the stirring head (collet) 502 increases when the plungerdescends. This end is sized to fit loosely but closely within theannular wall 47 at the top of stirrer 40 when the collet is notexpanded. When plunger 526 descends, the fingers 504 expand outwardly towedge against the inside of wall 47, in manifold M, securely engagingthe stirrer.

In operation, the stirring head 502 is first lowered so that the colletenters the manifold M. The dashed motor and bracket lines in FIGS. 33and 34 indicate this lowered position. Then plunger 526 descends to lockthe stirring head to the stirrer. Then the stepper motor 514 is operatedto slightly raise the stirring head and the attached stirrer 40. Thisvertical movement need only be very small, such as 0.050 in., just tofree the stirrer from the ribs 26 and prevent interference with thecontainer during stirring. Then DC stirring motor 518 is operated inaccordance with the specimen-specific stirring protocol. Stirring speedcan vary, and is usually in the range of about 500 rpm to about 3,000rpm. The stirring time can vary from about 5 seconds to about 90seconds. The base or bottom wall 41 of the stirrer acts as a slinger tothrust any liquid that may rise along the stirrer against the containerwall, and prevents the escape of liquid from the container. Withdrawingthe plunger 526 from the collet releases the stirrer 40 from the collet502 so the specimen container can move on to the next station.

A contracting collet could be used instead of expanding collet 502. Inthat case, the collet fingers would fit around the outside of annularwall 47, and would be squeezed together to clamp around the wall by adescending sleeve that surrounds the fingers.

Filter Placement Station

At the filter placement station 600 an appropriate filter assembly F(see FIG. 5) is loaded into the open manifold M at the top of thestirrer 40. Filter assemblies can come in different filterconfigurations for automated machine recognition. For example, one setof filter assemblies can be colored red (5 micrometers), another setwhite (8 micrometers), each having different filtering properties, and acolor sensor can detect which type of filter is before it and cause theproper filter to be loaded. The filter assemblies are dispensed by apusher from a magazine having multiple filter tubes.

FIGS. 36-40 show the structure and operation of the filter placementstation. Referring to FIGS. 37 and 40, a filter dispensing head 610comprises a filter magazine in the form of a turret 612 rotatable on aspindle 614 by a stepper motor 616. Vertical post 611 provides the mainsupport for the turret. Turret 612 has a top support plate 618 witheight equally spaced holes 620 near its periphery, each hole openingthrough the edge of the plate 618 with a slot 622. A bottom guide plate624 on spindle 614 has a similar arrangement of holes that are alignedwith the holes and slots in the top support plate.

Eight steel filter tubes 626, each having an upper support shoulder 628,are supported vertically in holes 620 and the aligned holes beneaththem, with shoulders 628 resting on the top of top plate 618. Eachfilter tube 626 has a full-length slot 630, and its bottom portion issplit into four springy fingers 632 by slots 634. Just above the bottomend the fingers 632 curve inwardly, forming rounded inner shoulders 636against which a filter assembly F rests. The filter tube is dimensionedsuch that the shoulders 636 keep up to a full stack of filter assembliesF from falling out of the tube, but deflect to allow a filter assemblyto pass when the stack is pushed downwardly without damage to the filterassembly. Fingers 632 thus form a springy choke.

FIG. 39 shows the position of the filter magazine 612 in relation to theprocessing path and the adjacent processing stations, namely the primarystirring station 500 to the left, and the specimen acquisition station700 to the right, all located on one side of the processing path asdefined by guide rails 250. On the other side of the processing pathopposite the filter magazine 612 is the assembly that supports anddrives a pusher arm 640. This assembly comprises a support post 642supporting a Z-axis lead screw 644 driven by a stepper motor (not shown)which moves a shuttle 646 that carries pusher arm 640. A filter sensor650 positioned opposite bottom guide plate 624 monitors the passage(drop) of the lowest filter assembly F in the filter tube presented to(i.e., directly above) the specimen container. Sensor 650 also detectswhen the filter tube is empty. A second sensor 651 monitors filter type.

Filter assemblies of the same type are stacked in the properorientation, with the membrane filter side (beveled edge) facing down,in each tube. For example, 54 filter assemblies can be housed in eachtube; thus a total of 432 filter assemblies can be loaded into themagazine. Fifty-four filter assemblies can be prepackaged in a stackthat is inserted into a filter tube with a wrapper tab projecting fromslot 630, and unwrapped by pulling the tab outwardly. Alternatively,filter assemblies of the same type can be dumped onto a vibratoryfeeder, which can recognize their orientation by geometricconfiguration, and properly orient and feed the filter assemblies ontothe tubes. Several of these feeders can be used, one for each type offilter assembly.

In operation, with the pusher arm 640 in its home (top) position,indicated by the dashed shuttle outline in FIG. 38, the filter magazine612 is rotated by stepper motor 616 until sensor 650 detects thepresence of the specified type of filter assembly in the filter tubebefore it. Shuttle 646 then moves downwardly with pusher arm 640 movingthrough slot 630 to press the stack of filter assemblies in that tubedownwardly, until the lowest filter assembly drops from the tube intothe manifold M in stirrer 40. When filter drop is sensed, the shuttle646 with its pusher arm 640 stops its advance. In an alternativearrangement, a weight sensor can be used to monitor the weight of thefilter stack, and detect by weight change when a filter assembly hasdropped from the stack and when the filter tube is empty.

The use of eight filter tubes 626 in magazine 612 enables unattendedprocessing of all of the specimens housed in the trays of the vialautoloader 300. For a counter-top model of the type described above,however, a single filter tube supported in a fixed position above theprocessing path would suffice for processing specimens that require thesame type of filter.

Specimen Acquisition and Cell Deposition Station

Referring to FIG. 41, specimen acquisition station 700 has a suctionhead 702 that descends to engage the upper portion of the stirrer 40.Before drawing a vacuum on the specimen through the filter assembly F,the suction head grips, slightly lifts and rotates the stirrer 40, thistime more slowly than at the primary stirring station (typically no morethan 500 rpm for a 5 second interval), to re-suspend the particulatematter in the specimen liquid. The re-stir motor can be a Maxon 24 voltDC planetary gear-reduced type. Then suction is applied through suctionline 750 to aspirate specimen liquid from the container 20 throughsuction tube 43, into the particulate matter separation chamber(manifold) 46 and through the filter assembly F, leaving a monolayer orthin layer of uniformly deposited cells on the bottom surface of thefilter as described above. It may also be possible to rotate the stirrerslowly while the specimen liquid is being aspirated.

FIG. 6 shows how the suction head cooperates with the annular wall 47 ofthe stirrer manifold and the filter assembly F therein. The outerportion 704 of the suction head envelops the wall 47 and has an O-ring760 that seals against the outside of wall 47. The inner portion 706 ofthe suction head has two concentric O-rings 762, 764 that seal againstthe top of filter holder 200. Suction applied through port 750 creates avacuum around central opening 204 and within filter holder 200, whichdraws liquid into the manifold 46 and through the filter 202. An O-ring766 is interposed between the inner and outer portions of the suctionhead.

Referring to FIG. 42, when aspiration of the specimen is complete, thesuction head 702 is raised. The inner portion 706 of the suction head isextended at the same time by action of a pneumatic cylinder (not shown)mounted above the suction head. As the suction head 702 is raised, theouter portion 704 disengages from the stirrer 40, but the filterassembly F is retained on the inner portion 706 by application of avacuum through suction line 752 to the annular space between O-rings 762and 764. Thus the suction head 702 removes filter assembly F from thestirrer, and can continue to apply light suction via suction line 750through the filter to effect a desired degree of moisture control of thecellular material on the filter.

The suction head 702 then moves laterally away from the transportconveyor by pivoting 90° about a vertical axis to the cell transferposition “P” shown in FIG. 46, to position the filter assembly F over amicroscope slide S delivered from a slide cassette at slide presentationstation 900. This pivoting movement of suction head 702 can also be seenin FIGS. 11 and 39. The inner portion 706 of the suction head 702 thenmoves downwardly to press the filter against the slide S with a tampingforce in the range of 4 to 8 lbs. and transfer the monolayer of cellsthereto. The phantom lines in FIG. 42 show this change in position ofsuction head 702 and contact of the filter with slide S. Instead ofbeing pivotally mounted, the suction head 702 could be mounted forrectilinear movement to and from a different deposition site whereslides are presented, e.g., above the processing path.

Referring to FIGS. 43-46, suction head 702 is rotatably mounted on aboom 716 that also carries the re-stirring motor 718, which rotatessuction head 702 through a timing belt 720. Boom 716 is pivotallysupported about a vertical axis 721 on a slide 722, which is verticallymovable along frame support 724 by means of a Z-axis stepper motor 726and a lead screw 728. Motor 726 thus moves the entire suction headvertically. Pivoting motion of boom 716 is effected by stepper a motor717 operating through a gear train (not shown). Vertical motion of theinner portion 706 of the suction head is effected by a pneumaticcylinder and return spring (not shown) mounted above the suction head toan L-bracket 719, substantially identical to the arrangement 412, 413,415 (see FIG. 29) used to move the plunger 408 of the uncapping head402.

The frame support 724 is mounted on a slide 730 so as to be movablelaterally of the transport path. A Y-axis stepper motor 732 and a leadscrew 734 effect this movement. After the slide is printed the suctionhead is raised by the Z-axis motor, and the Y-axis stepper motor 732advances the entire assembly to the dashed line position “X” shown inFIG. 43. Then the suction head pivots back to its original orientation,transverse to the transport path (position “S” in FIG. 46). The Y-axisstepper motor 732 then pulls the entire assembly back toward itsoriginal position (solid lines in FIG. 43). As the suction head 702moves (to the right as seen in FIG. 43), the still-retained filterassembly F is “scraped” off the suction head by the edge 736 of anopen-top used filter (waste) tube 738 (see also FIGS. 11 and 39). Thisleaves suction head 702 free to engage a fresh filter assembly.

The vacuum source that communicates with the suction head 702 pulls aslight vacuum, e.g., in the range of 3 in. to 10 in. Hg (adjustable by aregulator), through suction line 750 to aspirate specimen liquid anddraw it through the filter assembly F. The separately regulated vacuumapplied through suction line 752 for holding the filter assembly to thesuction head 702 is higher, on the order of 20 in. Hg.

Formation of high-quality specimens on microscope slides dependscritically on the deposition of a monolayer of cells of specifiedconcentration (i.e., number of cells per unit area) on the surface ofthe filter that will contact the slide. That, in turn, dependscritically on the aspiration rate and/or the aspirated flow volume.Since cell concentration on the filter surface is a function of thenumber of filter pores blocked by the solids suspended in the specimenliquid, the percent of flow reduction from the maximum open filtercondition correlates to the blockage or amount of accumulation on thefilter. Because of the nature of biological specimens, solid particleconcentration is a significant variable in the process and must be takeninto consideration. Also, it is important to identify the total volumeof material filtered on a real time basis for other processingoperations.

The specimen acquisition station thus further includes a depositioncontrol system for controlling the liquid draw vacuum duration bymonitoring the flow rate and/or aspirated volume. The monitored flowrate or aspirated volume can be used to signal vacuum cut-off and/orsuction head retraction, which correlates to the specified concentrationof cells collected on the membrane filter surface. If a specifiedconcentration factor is not achieved before a specified volume of fluidis aspirated, the system can also issue a retract signal.

Different types of deposition control systems or modules can be used forthese purposes. FIG. 47 schematically shows one such system, which has ameter in the form of a digital level detector positioned along a fluidcolumn. This “bubble flow” system can use sensors in the form of aplurality of LED emitters and corresponding number of photosensors, suchas Omron sensor, EE-SPX613 GaAs infrared LED, placed along the length ofthe column. Any other type of sensors may be used. Alternatively, LEDsensors such as the Omron sensors mentioned above can be used withoutcorresponding emitters when they are positioned just at the edge of aglass tube. The meniscus edge of the liquid in the tube diffracts thelight passing through the tube, and the sensor will detect the shiftedlight pattern when the rising meniscus edge reaches the sensor.

The fluid column is formed in a vertically extending transparent tube orcylinder 770, e.g., one made of Pyrex glass 9 mm in diameter by 1 mmthick. The aspirated specimen fluid is drawn from the specimen containerthrough the membrane filter, and pulled into the glass cylinder 770 viasuction line 750 and a 3-way valve 778, by means of a vacuum source 772connected to the top of the cylinder. The sensors 774 are positionedevenly along the length of the cylinder 770, preferably at 1.5 mlcapacity intervals, and are interfaced with a controller ormicroprocessor 776.

In operation, in the normal state, with no fluid in the tube 770, thesensor relay line is “low.” Vacuum begins to draw fluid into the tubethrough the filter, and the controller marks the beginning of the drawsequence. When the fluid reaches the first sensor, the first sensorrelay line goes “high.” The controller marks the time it took for thefluid to reach the first sensor, indicating the nearly free-flowcondition of the filter, and the relative viscosity of the fluid in thetest. When an additional 1.5 ml of fluid is drawn into the tube, thesecond sensor relay line goes “high.” The time interval for the first1.5 ml of fluid (between the first and second sensors) is noted by thecontroller, and this becomes the reference time base. As each additional1.5 ml of fluid is drawn into the system (and is detected by succeedingsensors), the time base for that increment is computed. When theincremental time base reaches an empirically derived percentage (e.g.,120%) of the original (reference) time base, the controller indicatesthat cell collection is completed, and a stop signal is transmitted,preferably to retract the suction head 702 from the manifold in thespecimen container. The empirically derived figure mentioned above isvariable with the protocol and directly controls the cellularity of thespecimen sample.

The best approximation of the free-flow condition of the filter isobtained if the time it takes for the fluid to reach the first sensor774 is kept to a practical minimum. This can be accomplished byincorporating the first sensor into the suction head itself, asschematically illustrated in FIG. 47 a. In this embodiment, innerportion 706 of the suction head carries an emitter 774 a and an opposedsensor 774 b, which detects the leading edge of the fluid column veryclose to the filter assembly F. The outer portion 704, which has teeth775 engaged by timing belt 720 (not shown), is rotatable about the innerportion 706 (note interposed bearing 773) to rotate the stirrer (notshown) and stir the specimen prior to aspiration.

During the specimen drawing operation, the controller records thecumulative or total aspirated volume. If the cumulative volume reaches apredetermined level before reaching the predetermined flow ratereduction from the reference flow, the controller will also issue a stopsignal and a flag indicating that the stop signal issued not as a resultof desired reduced flow, but by reaching the maximum liquid draw limit.A slide formed under the flagged condition will likely form ahypo-cellular condition. The controller can imprint the slide andindicate to the DMS that a hypo-cellular condition likely exists.Accordingly, if the flagged condition exists, the controller issues asignal to purge the liquid in the cylinder 770 and initiate a seconddraw. The cylinder is purged of all liquid after each sample is taken.

Referring to FIG. 48, the deposition control system can have a purgevalue so that when the draw cycle is completed, the stop signalgenerated by the controller 776 will open the purge valve to vent thevacuum supply line to the atmosphere and divert the liquid remaining inthe cylinder 770 into a waste container. The cylinder 770 can bemaintained under a negative pressure. The system is then ready for thenext cycle. Specifically, the system can have a 2-way solenoid valve V-3in the suction line with one port 780 open to the atmosphere. The bottomof the cylinder 770 is connected to a valve manifold 782 with twosolenoid valves V-2, V-4. The solenoid valves can be Lee LF seriesdesigned for use in vacuum systems, 2-way valve LFVA 2450110H, vitonseal, 24 volt and 3-way valve, LFRX 0500300B, viton seal, 24 volt. The2-way valve V-4 can port the specimen liquid to the bubble flow cylinder770, or to vacuum by-pass 784. The 2-way valve V-2 can control thefilter dehydration vacuum source. FIG. 49 illustrates the valve logic.

The deposition control system can use an analog level indicator insteadof the digital sensors 774. The analog level indicator sensescapacitance of the aspirated liquid. The difference is only in themethod of sensing the volume and fill rate of the liquid in the cylinder770. Here two spaced electrodes are used, one around the outside of thecylinder 770 and the other positioned down the center of the cylinderthe cylinder, separated from the aspirated liquid by a dielectric. Ahigh frequency, such as 10 kHz, low voltage current is applied acrossthe electrodes. Capacitance in this system is measured by a bridgecircuit, which provides an analog indication of capacitance in thecircuit. As fluid fills the column, capacitance in the circuitincreases. A 10×differential in direct capacitance is easily obtainedwith this system. Capacitance is indicated on a real time basis and canbe sampled frequently enough to provide control of the sampling system.This arrangement, like the first two, uses a computer or microprocessorand a bubble flow technology to measure the flow rate and the totalfluid volume in real time. The predetermined volume increment for thesearrangements can be in the range of about 0.1 ml to 5.0 ml, andpreferably is in the range of about 1.0 to 2.0 ml.

A different system can use an ultrasonic indicator for measuring fluidmovement through a tube. The ultrasonic system uses ultrasonic wavepropagation through a moving liquid. In this regard, the third systememploys an ultrasonic emitter and detector clamped across the liquiddraw tube (suction line 750) operating on the distal end of the filterassembly F. This system provides a digital indication of fluid flow inthe tube, the total volume aspirated through the tube being calculatedby a flow interval calculation. It measures phase shift from theultrasonic wave generator source to a detector for measuring flow speed.

Another way to measure aspirated fluid volume and control the durationof the specimen draw is to detect the change in the weight of thespecimen vial. This can be accomplished by using a sensor that makes ahigh-precision measurement of the weight or mass of the vial containingthe specimen that is being aspirated. Vial weight or mass is repeatedlymeasured at a high frequency such that the rate of change of the weightor mass of the vial is accurately determined. Specimen aspiration iscompleted when the rate of change in weight or mass has diminished by apredetermined amount or percentage from the initial rate. The weightsensor can be, e.g., a load cell in each conveyor receptacle 246, or asingle load cell beneath the conveyor at the specimen acquisition headthat rises to engage the container above it. In either case, thespecimen acquisition head can be raised slightly during aspiration tounload the container so that the load cell can measure only the combinedweight of the container and the remaining specimen.

Although specimen acquisition preferably is accomplished throughaspiration (using a vacuum), it can also be accomplished by pressurizingthe container 20 through an appropriate head that seals against the topof the container and forces specimen liquid up through tube 43 andthrough the filter assembly by means of positive pneumatic pressure. Thefluid volume control schemes and mechanisms described above would alsowork in conjunction with such a pressurized specimen acquisition system.

The cell concentration can be selected from low to high by defining flowcontrol cut-off. For a typical low cellularity result, the cut-off canbe 80% of the 120% reference discussed above, and for high cellularitythe cut-off can be set at 60% of the reference, selectable in 5%increments. The number of slides per specimen can range from one tothree. Some of the typical default protocols are as follows:

-   -   GYN: 1,000 RPM stir, 30 second interval, 8-micrometer filter,        60%—high cellularity, one slide.    -   Urine: 1,000 RPM stir, 20 second interval, 5-micrometer filter,        70%—medium cellularity, one slide.    -   Lung sputum: 3,000 RPM stir, 120 second interval, 5-micrometer        filter, 80%—high cellularity, two slides.        Re-Capping Station

After completing the specimen processing cycle, the specimen containeris resealed with the stirrer still inside the container. It is preferredto use a thin, polypropylene-coated aluminum foil to form the new cap,which is available in roll form. The foil is drawn across the open endof the specimen container, thermally bonded to the container at a sealtemperature of about 365° F. applied for about 3 seconds with a sealforce of 3 pounds, and cut from the roll. Of course, any other type ofre-capping material can be used as long as it is compatible with thevial material and creates a safe and reliable seal. For example, a foilbacked with a thermosetting resin adhesive could be used; asticky-backed foil could be used that does not require heat to effect aseal; or a plastic seal material can be bonded to the containerultrasonically. To enhance unattended operation, an automatic threadercould be included for threading a new roll of sealing material into there-capping mechanism. Cutting caps from a roll can be eliminated ifroll-mounted pre-die-cut closures having peel-off tabs are fed to there-capping mechanism.

Referring to FIGS. 50 and 52, the re-capping mechanism 800 has a sidesupport plate 802 secured to the machine base plate. The side supportplate carries a main frame 810 having a top plate 812 with slots 814,816, and two side plates 818, 820. A driver capstan 822 is journaled inside plates 818, 820. A foil advance motor 824, mounted on a bracket826, drives the capstan. A pressure roller 828 is pivotally mounted tothe main frame 810 and resiliently engages the capstan under theinfluence of a spring 830. Capstan 822 and pressure roller 828 definebetween them a throat through which the foil runs, and have resilientsurfaces which grip the foil for positive feed. A handle 832 allows thethroat to be opened manually to allow the end of the foil to be fed intothe throat after first passing through slot 814. A spindle 804, carriedside support plate 802, supports a replaceable roll of foil.

FIG. 51 shows the foil path 834 through the throat. An L-shaped cutter836 is pivoted at its elbow to the rear of main frame 810. One end of asingle-acting pneumatic cutter actuator cylinder 838 is mounted on abracket 840, and the other end of the cylinder is linked to the upperleg 842 of cutter 836. The lower leg of the cutter has a blade 844 thatnormally rests above the foil path downstream of the throat, held inthat position by a spring 845 linked between the upper leg 842 and thesupport plate 802.

A rear post 850 pivotally supports an arm 852 that extends forwardlytoward main frame 810. Arm 852 carries a heated platen 854 and a foilguide fork 856 having two tines that extend toward the throat and arespaced apart so as to allow the platen 854 to pass between them. Arm 852is kept elevated, in the rest position shown in FIG. 51, by a spring858. During the re-capping operation a single-acting pneumatic cylinder860 pulls down on the arm 852 to lower the platen 854 and the guide fork856. Note the position of a container 20 in a transport receptacle (notshown) beneath the platen 854.

In operation, the foil advance motor turns the capstan 822 to feed ameasured length of foil past the cutter blade 844, into the fork 856,and to the position shown by the dashed line in FIG. 51. A photocell 862detects the leading edge of the foil and signals the motor to stop. Thencylinder 838 is actuated to cut the foil, and cylinder 860 is actuatedto pull arm 852 down to the seal position. The cut length of foil issandwiched between the platen 854 and the container 20, and thecontainer is sealed. After about three seconds cylinder 860 isdeactivated and the arm 852 rises, returning to its rest position. Avacuum assist (not shown) optionally may be used to help hold the cutlength of foil in position on the platen prior to sealing.

The foil caps applied by the re-capping mechanism are approximatelysquare in shape. The corners of the foil caps can protrude from thevials and interfere with other re-capped vials that are returned to thetrays 330. Accordingly, a foil folding ring 870 (seen in phantom linesin FIG. 51) preferably is provided which acts to fold the edges andcorners of each foil cap down along the side of the container. The foilfolding ring 870 preferably is mounted to act on the vial in thetransport position immediately downstream of the re-capping mechanism,i.e., position “FF” in FIG. 51, and may be mounted on the recappingmechanism itself, e.g., to main frame 810, so that actuation of cylinder860 serves simultaneously to apply a foil cap to one container and foldthe edges and corners of the foil cap of the preceding (downstream)container. Alternatively, the foil folding ring or an equivalent foilfolding mechanism can be mounted further downstream of the re-cappingmechanism so as to act independently thereof.

Foil folding ring 870 is a metal ring having an inner diameter that isslightly larger than the outside diameter of the threaded portion of thecontainer 20. The ring 870 is mounted on an arm (not shown) that movesdownwardly when actuated to lower the ring 870 over the upper end of thecontainer. As the ring encircles the container, it folds the overhangingportions 872 of the foil cap against the side of the container. When thering rises after folding the foil, the container is held in position inits transport receptacle by a pin (not shown) that is mounted on a leafspring (not shown) and is situated in the center of the ring 870. Theleaf spring is carried by the arm that holds the ring, so the pinresiliently presses down against the center of the foil cap until thearm and the ring retract fully.

The foil seals applied to the processed containers are easily puncturedby a syringe or a pipette to obtain further liquid specimen samples. Theseals are very durable, however, withstanding rough handling andpreventing leakage in low ambient pressure conditions, e.g., in aircraftflying as high as 40,000 ft. Further, the appearance of the foil sealmakes it readily distinguishable from the cover of an unprocessed vial,making handling by low-skilled operators virtually foolproof. To avoidthe potential of puncturing the foil seal inadvertently, the re-sealedcontainer can be capped with an unused screw-on cover of a distinctcolor.

Slide Handling and Presentation

The LBP device can use 30 and 40 slide plastic magazines (cassettes),which can accept standard 25 mm×75 mm×1 mm and 1×3×0.040 in. slides.Metric and inch based slides can be used interchangeably. FIGS. 52-55show a 40-slide cassette C suitable for use in the LBP device. The slidecassette is in some respects similar to that disclosed in U.S. Pat. No.5,690,892 (incorporated herein by reference), but is specially adaptedfor use in other devices as well, such as an automated stainer, anautomated image analyzer, and a pathology work station, so that theslides do not have to be unloaded and reloaded into different magazinesfor use in those devices. Machine-readable indicia on the cassette, suchas a bar code or an embedded microchip, provides cassette informationthat can be linked by the DMS to the bar codes on the slides in thecassette so that the location and status of any cassette and any slidein that cassette can be tracked in a laboratory system. The cassettesare stackable for compact storage and easy retrieval.

Specifically, the slide cassette is molded of plastic and has agenerally rectangular shape with an open front 902, a rear wall 904, atop wall 906, a bottom wall 908 and side walls 910. The top wall 906bears bar-coded information 909. A guide flange 912 extends laterallyoutwardly from each side wall. Rear wall 904 has a rectangular centralopening 914 through which a slide shuttle can pass (see below) toextract and return one slide at a time. An inwardly projecting ridge 916around the central opening acts as a stop against which the slides abutwhen they are inserted into the cassette. The preferred material for thecassette is ABS plastic; alternative choices include polyurethane,thermoplastic polyester, and polypropylene. The open front face is sizedto accommodate the rear of another like cassette so as to be stackable.

The slides are supported on shelves 918 at each side of the cassette. Inthe illustrated embodiment there are 41 pairs of left and right shelves,and each pair (except for the top pair) supports one slide that spansthe space between the shelves. Referring to the detailed view in FIG.53, each shelf (except for the top and bottom shelves) has a raised topledge 920 on which the slide rests and an underside beam spring 922 forapplying a force to pinch and thereby frictionally restrain the slideagainst the top ledge directly beneath it. This arrangement keeps theslides from falling out of the cassette, even when the cassette is heldface down, yet enables each slide to be moved out of and back into thecassette by the slide presentation apparatus, described below, withoutblocking, scratching or interfering with the slide-mounted specimens.Each shelf 918 also has a lead-in ramp 924 which guides the slide duringinsertion into the cassette. Each shelf 918 (including spring 922)preferably is integrally molded into the cassette and is attached toboth the rear wall 904 and a side wall 910. However, separatelyfabricated springs, plastic or metal, may be inserted between theshelves instead.

Each side wall is provided with multiple drainage ports 926 which allowfluid to drain from the cassette after removal from a staining bath. Thelast (top and bottom) drainage ports 923 on each side also cooperatewith a hanger assembly of a stainer for moving the cassette from onestaining bath to another. During the staining operation the cassette isoriented generally on its side, hung from the last two drainage ports onthe upper side. An all-plastic construction makes the cassettecompatible with acid baths and all types of staining bath compositions.

Referring to FIG. 54, rear wall 904 has two rows of apertures 927 thatform two integrally molded gear racks 928, which are adapted to engagepinion gears 936 (see below) for moving the cassette longitudinally sothat each slide can be accessed by the slide shuttle. Two spacedparallel racks and two pinion gears enhance the smoothness and accuratepositioning of the cassette, as compared to a single rack and singlepinion. Also integral with the rear wall is a row of 40 cassetteposition sensing slots 929 extending through the rear wall andcoincident with the positions of the slides to allow for optical sensingof each slide. Further, rear wall 904 has a row of 40 blind recesses 925(these do not extend completely through the rear wall) that allow foraccurate sensing of cassette position when it is driven via the gearracks 928.

The molded cassette preferably is supplied wrapped in sealed plastic forcleanliness, with slides installed. It is therefore well suited forshipping, relatively low in cost, disposable yet reusable. It has a highstorage capacity and is stackable with others, thus providing highdensity storage for specimen samples.

Slide cassettes populated with slides are manually loaded into the LBPdevice in an elevated in-feed track 930 (see FIG. 11) located behind thefilter loading station 600 and the specimen acquisition station 700. Nolatching is required to enter cassettes into the system. Up to tenunprocessed cassettes can be loaded in the LBP device at any one time,but only in a single orientation. The cassettes can be marked with a topindicator, and will not be accepted if they are installed backwards orupside down. The cassettes are loaded with their open fronts facing tothe right as seen in FIG. 11, with the lead cassette between verticalrails 932.

The lead cassette moves down incrementally whenever a new slide is to bewithdrawn from the cassette for specimen printing. This is accomplishedby a stepper motor (not shown) driving pinion gears 936 that engages theracks 928 on the back of the cassette C (see FIG. 54). When all slidesin the cassette have been processed, the cassette descends all the wayto outfeed track 940, and a stepper motor/lead screw pusher 938 movesthe cassette to the right into outfeed track 940, and then retracts.Then the next cassette in the infeed track 930 is advanced by amotor/lead screw pusher (not shown) to the front position betweenvertical rails 932, where it is engaged by the pinion gears 936 andmoved downwardly until the first (lowest) slide comes into position forextraction. Each of the feed tracks can have a home sensor, which can beOmron self-contained shutter type, and a cassette full sensor, which canbe Keyence fiber optic.

FIGS. 11, 56 and 57 show the slide presentation system, which uses aslide shuttle feed system 960, e.g. AM Part No. 5000-1, for extractingone slide at a time from the cassette along the X-axis and placing it ona Y-axis handler, which moves the slide to the pressing (print)position. The aforementioned U.S. Pat. No. 5,690,892 discloses a similarslide cassette and shuttle arrangement used in a pathology work station(microscope). The Y-axis handler 962 has a slide platen 964 secured to afollower 966, 967. The handler is driven by a stepper motor 970 and alead screw 972, guided along a rail 968. A slide is held to the platenunder a fixed shoulder 974 (against a spring 976) and a pivoted arm 978which is spring-biased in the counterclockwise direction as seen in FIG.56.

When the handler 962 moves to the left, arm 978 moves off an adjustablestop 980 and rotates over the slide. The full Y-axis slide travel (shownas “T” in FIG. 57) brings the center of the slide to the print position“P” (note the dashed line position of the slide and the handler in FIG.56). On its way to the print position the bar code number on the slideis acquired by a bar code reader 982 and transmitted to the host database. When the print position is reached the suction head 702, which haspivoted along arc “A” about axis 721, lowers the filter assembly F intocontact with the slide, as described above, depositing (printing) thespecimen on the slide. Vacuum on the filter is maintained throughout theprinting cycle to prevent over-hydration of the sample and unintentionaldripping.

After printing the slide moves back to the right, pausing under afixative dispensing head 984. Here a solenoid-driven pump (not shown),such as Lee LPL X 050AA, 24V, 20 microliter per pulse, yielding 12microliters per pulse (maximum of 2 pulse/second), applies fixative tothe specimen. The total volume can be determined by the number ofsolenoid cycles. The total fixative volume dispensed is programmable in20 microliter increments. It can have a flexible connection to adispensing sapphire jet nozzle with a 0.030 in. orifice. The liquid canbe gravity-fed from a reservoir to the pump. The reservoir can be a tankand can have a “fluid low” sensor connected to the operating system.More than one fixative dispenser can be employed to provide alternativefixatives as determined by processing protocols.

After the specimen is fixed, the completed slide moves all the way tothe right, where it is transferred by the slide shuttle mechanism backto its original position in the cassette. When the cassette is fullyprocessed, the entire cassette is ejected into the outfeed track 940, asdescribed above.

A Complete Laboratory System

The present LBP device does not require that specimens be pre-processedbefore loading, and can automate every step of the slide preparationprocess. Moreover, the device does not require the operator to open anyof the specimen containers—an important operator safety feature. The LBPdevice can automatically prepare high quality cytology slides from allspecimen types, including mucous-containing GYN and non-GYN specimens,using the integral high-speed, high-shear mixing station thatfacilitates mucous disaggregation. The incorporated dual-flow filtersystem allows production of slides with optimal cell separation, cellconcentration, cell dispersion, and optimal preservation of antigens,DNA, and morphologic characteristics to enhance the performance ofsubsequent testing. The slide cassettes, containing up to 40 slideseach, will be utilized in the follow-on laboratory processing devices toavoid the labor-intensive need to transfer slides to different racksbefore continuing with slide processing. Data on the patient, thespecimen, the vial, the cassette and the slide can be transferredautomatically to the LIS over the user's network, via a DMS softwareinterface.

The present LBP device can provide eight hours of unattended operation.Thus, if the operator re-loads the device before leaving for the day, asingle-shift laboratory can produce two shifts of output per day withoutadded personnel or equipment costs. The total throughput can exceed160,000 slides per year, at a per-test cost significantly below that ofthe current leading LBP system.

The LBP device also has the capability to process specimens for currentand future molecular diagnostic tests including quantitative DNAanalyses, and tests utilizing markers & probes. Features built into thedevice include the capacity to employ multiple fixative dispensers inorder to provide non-routine fixatives that may be required for specialmolecular diagnostic tests.

The complete laboratory system, illustrated, e.g., in FIG. 21 a,includes a pathology review station, a computer-aided microscopy workstation used by pathologists to review specimen slides and sign outcytology cases. As with all components of the laboratory system, thepathology review stations are networked to the DMS and thereby to allother devices on the system, for rapid access to patient data andspecimen processing information. The pathology review station acceptsslide cassettes for automated loading and review of specimen slides.Computerized, fully automated image analyzers will perform quantitativeanalyses of DNA and molecular diagnostic tests, receiving theiroperating instructions and reporting their results via specimen barcodes using the integral DMS. See, for example, AccuMed/MDI U.S. Pat.Nos. 5,963,368; 6,091,842; and 6,148,096, which are incorporated hereinby reference.

The laboratory system will also include, for example, slide autostainersand autocoverslippers (and/or combination automated stainer/coverslipperdevices) controlled via the DMS that utilize the same slide cassette asthe present LBP device. Cassettes containing processed slides can beutilized directly in these additional devices without the need to unloadslides and reload them into separate racks.

The inter-connectivity and high degree of automation of the processingand analytical devices making up the laboratory system will enablehigh-quality, high-throughput specimen processing and analysis atrelatively low cost.

INDUSTRIAL APPLICABILITY

The above disclosure presents a safe, effective, accurate, precise,reproducible, inexpensive, efficient, fast and convenient vial-basedsystem and method for collecting, handling and processing liquid-basedcellular specimens, providing fully integrated specimen and informationmanagement in a complete diagnostic cytology laboratory system.

1. An automated method for individually processing multiple specimens ofparticulate matter-containing liquid in respective containers, eachcontainer having a mixer therein, the method comprising: transportingthe containers in single file along a single processing path to presentthem seriatim to a single mixing head and thereafter to a singlespecimen acquisition head, the mixing head adapted to mix the specimenin any container presented to it, and the specimen acquisition headadapted to aspirate previously mixed particulate matter-containingliquid from any container presented to it and pass the aspirated liquidthrough a respective filter so as to collect a particulate matter sampleon a surface of the filter; actuating the mixing head in response topresentation of a container thereto and irrespective of the operationalstate of the specimen acquisition head so as to engage the mixer in thecontainer and carry out the mixing operation independently of theoperational state of the specimen acquisition head; and actuating thespecimen acquisition head in response to presentation of a containerthereto and irrespective of the operational state of the mixing head soas to carry out the aspirating operation independently of theoperational state of the mixing head.
 2. An automated method accordingto claim 1, wherein operation of the mixing head is carried out inaccordance with a protocol specific to the specimen presented thereto,and operation of the specimen acquisition head is carried out inaccordance with a protocol specific to the specimen presented thereto.3. An automated method according to claim 1 or claim 2, furthercomprising pressing seriatim each filter through which aspirated liquidhas passed against a respective slide positioned in proximity to theprocessing path to transfer the particulate matter sample from thefilter to the slide.
 4. An automated method according to claim 1 orclaim 2, wherein the specimen acquisition head is adapted to mix thespecimen in each container presented to it, the method includingactuating the specimen acquisition head to mix the specimen immediatelyprior to the aspirating operation.
 5. An automated method according toclaim 4, wherein the specimen acquisition head mixes the specimen moreslowly than the mixing head.
 6. An automated method according to claim5, further comprising pressing seriatim each filter through whichaspirated liquid has passed against a respective slide positioned inproximity to the processing path to transfer the particulate mattersample from the filter to the slide.
 7. An automated method forindividually processing multiple specimens of particulatematter-containing liquid in respective containers, each container havingtherein a mixer with an upper particulate matter separation chamberadapted to receive a filter and a depending aspiration tubecommunicating with the separation chamber, the method comprising:transporting the containers in single file along a single processingpath to present them seriatim to a plurality of operating heads spacedso that the operating heads can operate simultaneously on differentspecimens, the plurality of operating heads comprising: a single mixinghead adapted to engage and move the mixer in any container presented tothe mixing head to mix the specimen therein, a single filter loadinghead downstream of the mixing head having a filter magazine and adaptedto dispense a filter into any particulate matter separation chamberpresented to the filter loading head, and a single specimen acquisitionhead downstream of the filter loading head adapted to seal against theparticulate matter separation chamber presented and apply suctionthereto to aspirate previously mixed particulate matter-containingliquid from the container and pass the aspirated liquid through thefilter in the separation chamber so as to collect a particulate mattersample on a surface of the filter; and actuating each of the operatingheads in response to presentation of a container thereto andirrespective of the operational states of the other operating heads soas to carry out the respective operation of each operating headindependently of the operational states of the other operating heads. 8.An automated method according to claim 7, wherein operation of themixing head is carried out in accordance with a protocol specific to thespecimen presented thereto, and operation of the specimen acquisitionhead is carried out in accordance with a protocol specific to thespecimen presented thereto.
 9. An automated method according to claim 7or claim 8, further comprising pressing seriatim each filter throughwhich aspirated liquid has passed against a respective slide positionedin proximity to the processing path to transfer the particulate mattersample from the filter to the slide.
 10. An automated method accordingto claim 7, wherein the filter magazine of the filter loading head isadapted to store at least two types of filters, and the filter typedispensed into the particulate matter separation chamber is governed byinformation specific to the specimen presented.
 11. An automated methodaccording to claim 7, claim 8 or claim 10, wherein the specimenacquisition head is adapted to mix the specimen in each containerpresented to it, the method including actuating the specimen acquisitionhead to mix the specimen immediately prior to the aspirating operation.12. An automated method according to claim 11, wherein the specimenacquisition head mixes the specimen more slowly than the mixing head.13. An automated method according to claim 12, further comprisingpressing seriatim each filter through which aspirated liquid has passedagainst a respective slide positioned in proximity to the processingpath to transfer the particulate matter sample from the filter to theslide.
 14. An automated method according to claim 7, claim 8 or claim10, wherein each container has a removable cover and is loaded into theprocessing path with the cover in place, and said plurality of operatingheads further comprises an uncapping head along the processing pathupstream of the mixing head and adapted to grip and remove the coverfrom a container presented thereto.
 15. An automated method according toclaim 14, wherein said plurality of operating heads further comprises acontainer recapping head along the processing path downstream of thespecimen acquisition head and adapted to apply a sealing cap to anuncapped container presented thereto.
 16. An automated method accordingto claim 15, further comprising unloading processed and recappedcontainers from the processing path, and loading unprocessed containersinto the processing path in positions vacated by processed and recappedcontainers.
 17. An automated method according to claim 16, whereincontainer unloading and loading is carried out while movement of allcontainers along the processing path is arrested.
 18. An automatedmethod for individually processing multiple specimens of particulatematter-containing liquid in respective containers, each container havinga removable cover, the method comprising: transporting the containers insingle file along a single processing path to present them seriatim to aplurality of operating stations spaced so that the operating stationscan operate simultaneously on different specimens, the plurality ofoperating stations comprising, in sequence: a single uncapping stationat which the cover of a container presented thereto is removed, a singlemixing station at which the specimen in a container presented thereto ismixed, a single filter loading station at which a filter is placed intothe particulate matter separation chamber adjacent the top of acontainer presented thereto, and a single specimen acquisition stationat which the previously mixed particulate matter-containing liquid isaspirated from the container and passes through the filter so as tocollect a particulate matter sample on a surface of the filter; andactuating each of the operating stations in response to presentation ofa container thereto and irrespective of the operational states of theother operating stations so as to carry out the respective operation ofeach operating station independently of the operational states of theother operating stations.
 19. An automated method according to claim 18,further comprising a container recapping station, downstream of thespecimen acquisition station, at which a container presented thereto isrecapped.
 20. An automated method according to claim 18, wherein thespecimen acquisition station re-mixes the particulate matter-containingliquid before aspirating it from the container.
 21. An automated methodaccording to claim 18 or claim 20, wherein a slide presentation stationis located in proximity to the processing path and the specimenacquisition station, the method further comprising pressing seriatimeach filter through which aspirated liquid has passed against arespective slide to transfer the particulate matter sample from thefilter to the slide.
 22. An automated method according to claim 21,further comprising a container recapping station, downstream of thespecimen acquisition station, at which a container presented thereto isrecapped.
 23. An automated method for individually processing multiplefluid specimens of biological material previously collected inrespective containers, the method comprising: placing each containerwith its previously collected specimen on a conveyor, and moving theconveyor to transport the containers in single file along a singleprocessing path to present them seriatim to a single preprocessingapparatus and thereafter to a single specimen acquisition apparatus, thepreprocessing apparatus adapted to preprocess the specimen fluid in anycontainer presented to it, and the specimen acquisition apparatusadapted to remove preprocessed specimen fluid from any containerpresented to it for subsequent analytical testing or evaluation;actuating the preprocessing apparatus in response to presentation of acontainer thereto and irrespective of the operational state of thespecimen acquisition apparatus so as to carry out the preprocessingoperation independently of the specimen acquisition apparatus; andactuating the specimen acquisition apparatus in response to presentationof a container thereto and irrespective of the operational state of thepreprocessing apparatus so as to carry out the fluid removal operationindependently of the operational state of the preprocessing apparatus.24. An automated method according to claim 23, wherein the preprocessingapparatus disperses particulate components of the specimen fluid.
 25. Anautomated method according to claim 24, wherein the specimen acquisitionapparatus collects a cytology sample.
 26. An automated method accordingto claim 24, wherein the specimen acquisition apparatus filters theremoved specimen fluid.
 27. An automated method according to claim 26,wherein the specimen acquisition apparatus collects a cytology sample.28. An automated method according to claim 27, wherein the specimenacquisition apparatus places the cytology sample on a slide.
 29. Anautomated method according to claim 3, wherein the containers areallowed to advance along the processing path after the filter pressingand sample transfer operation has begun.
 30. An automated methodaccording to claim 6, wherein the containers are allowed to advancealong the processing path after the filter pressing and sample transferoperation has begun.
 31. An automated method according to claim 9,wherein the containers are allowed to advance along the processing pathafter the filter pressing and sample transfer operation has begun. 32.An automated method according to claim 13, wherein the containers areallowed to advance along the processing path after the filter pressingand sample transfer operation has begun.
 33. An automated methodaccording to claim 21, wherein the containers are allowed to advancealong the processing path after the filter pressing and sample transferoperation has begun.